Objective To explore the value of ovarian-adnexal reporting and data system(O-RADS)MRI score combined with tumor markers(CA125+HE4)in ovarian tumors.Methods Data from 223 patients with ovarian tumors confirmed by pathology were analyzed retrospectively,including 260 lesions.The Kappa test was used to assess the consistency of O-RADS MRI score between low and high seniority physician groups.The receiver operating characteristic(ROC)curve was drawn to analyze the diagnostic effi-cacy of O-RADS MRI score combined with tumor markers(CA125+HE4)in ovarian tumors.Results The Kappa value of the O-RADS MRI score between low and high seniority physician groups was 0.803[95%confidence interval(CI)0.746-0.860].The sensitivity based on O-RADS MRI score for distinguishing benign and malignant ovarian tumors was 0.957 and 0.989,the specificity was 0.784 and 0.820,the accuracy was 0.846 and 0.881,the positive predictive value was 0.712 and 0.754,and the negative pre-dictive value was 0.970 and 0.993,and the area under the curve(AUC)was 0.871 and 0.905,respectively in the low and high senior-ity physician groups.Combined with tumor markers(CA125+HE4),the sensitivity and negative predictive value of both low and high seniority physician groups were 1.000.Conclusion The O-RADS MRI score has high diagnostic efficacy and good repeatability in ovarian tumors.Combined with tumor markers CA125 and HE4,the O-RADS MRI score can further improve the diagnostic sen-sitivity.

Objective Based on the phosphatidylinositol kinase(PI3K)/protein kinase B(AKT)pathway and M2 polarization of macrophages,the effects of polydatin on malignant phenotype of osteosarcoma cells were investigated.Methods The macrophages were divided into M0 group,M2 group,polydatin group,and polydatin+PI3K pathway agonist 740Y-P group.The expression of arginase-1(Arg-1)and CD206 mRNAs and proteins in macrophages were detected by RT-qPCR and Western blot.Western blot was used to detect p-PI3K/PI3K and p-AKT/AKT protein expression in macrophages.The osteosarcoma cells MG-63 were divided into control group,control group(RPMI-1640 medium culture),CM-M0 group(M0 group M0 macrophage supernatant is collected for osteosarcoma cells),CM-M2 group(M2 group M2 macrophage supernatant is collected for osteosarcoma cells),CM-polydatin group(M2 macrophage supernatant in the polydatin group was collected on osteosarcoma cells),and CM-polydatin+740Y-P group(M2 macrophage supernatant was collected on osteosarcoma cells in the CM-polydatin+740Y-P group).Edu staining assay was used to detect the proliferation of MG-63 cells.Scratches and Transwell assays were used to detect the migration and invasion of MG-63 cells.Results Compared with the M0 group,the Arg-1,CD206,IL-10,p-PI3K/PI3K,p-AKT/AKT increased in the M2 group significantly(P<0.05).Compared with the M2 group,the Arg-1,CD206,IL-10,p-PI3K/PI3K,p-AKT/AKT decreased in the polydatin group significantly(P<0.05).Compared with the polydatin group,the Arg-1,CD206,IL-10,p-PI3K/PI3K,p-AKT/AKT increased in the polydatin+740Y-P group significantly(P<0.05).Compared with the CM-M0 group,the proportion of Edu-positive cells in the CM-M2 group increased significantly(P<0.05),and MG-63 cell migration and invasion rates increased significantly(P<0.05).Compared with the CM-M2 group,the proportion of Edu-positive cells in the CM-polydatin group decreased significantly(P<0.05),and MG-63 cell migration and invasion rates decreased significantly(P<0.05).Compared with the CM-polydatin group,the proportion of Edu-positive cells in the CM-polydatin+740Y-P group increased significantly(P<0.05),and MG-63 cell migration and invasion rates increased significantly(P<0.05).Conclusion Polygonum cuspidatum can inhibit the proliferation,migration and invasion of osteosarcoma cells by inhibiting the M2 polarization of macrophages,and its mechanism may be related to the inhibition of PI3K/AKT signaling pathway.

Objective To investigate the effect of resveratrol(RES)on the function of osteoarthritis synovioblast(OA-FLS)and its molecular mechanism.Methods The purchased primary synovial cells were cultured to the third generation,and OA-FLS cell model was induced by IL-1β.The activity of FLS cells was observed by CCK-8 assay,and the appropriate RES intervention concentration was selected.The cells were divided into Control group,OA-FLS group,Low-RES group(1μmol/L)and High-RES group(5μmol/L).The effects of RES on the migration and invasion of OA-FLS were detected by cell scratch assay and Transwell assay.The effect of RES on OA-FLS apoptosis was detected by flow cytometry.The effects of RES on the expression of inflammatory factors in OA-FLS and re-lated proteins in NF-κB signaling pathway were detected by Western blot.Results The results of CCK-8 experiment showed that the concentration of RES above 10μmol/L significantly inhibited the viability of FLS cells,and the concentration of RES above 1 μmol/L and 5μmol/L had no significant effect on the viability of FLS cells.Compared with the Control group,the migration and invasion ability of OA-FLS group was enhanced,while the apoptosis ability was weakened,and the migration,invasion and apoptosis ability of OA-FLS group was significantly reversed after RES intervention.Western blot results showed that,compared with the Control group,the levels of inflammatory cytokines in OA-FLS group were increased,and the NF-κB signaling pathway was activated,while the treatment of OA-FLS with different concentrations of RES could reduce the expression of inflammatory cytokines and inhibit the transcription of nuclear fac-tors.Conclusion RES inhibited migration and invasion of OA-FLS,promoted apoptosis,and decreased the expression of inflammatory factors of OA-FLS,which may be related to the inhibition of NF-κB signaling pathway by RES.

From the perspective of new quality productivity,it is a new direction and opportunity to train innovative talents with modern technology and professional knowledge.This paper introduces the practice of"Topic-based"experimental teaching in the course of Clinical Immunology Laboratory Technology,and explores the possible ways to train new qualified personnel of medical labo-ratory technology.

Acupuncture and moxibustion therapy is a kind of treatment and health care method with original advantages of China.With the rapid development of epigenetics and systems biology technology,non-coding RNA(ncRNA)related research has made continuous breakthroughs in the field of acupuncture and moxibustion.This article collected the basic research literature on acupuncture and moxibustion related to ncRNA,and reviewed the research subsystems related to microRNA(miRNA),long chain non coding RNA(lncRNA)and circular RNA(circRNA).NcRNAs are widely involved in the growth,development and reproduction of the organism,as well as in the occurrence and development of various diseases,which fits with the multi-layer,multi-pathway and multi-target action network of acupuncture and moxibustion therapy.Taking ncRNAs as the breakthrough point to explore the mechanism of acupuncture and moxibustion in depth is not only conducive to promoting the exploration of new targets of acupuncture and moxibustion effect,but also can reveal the epigenetic regulation axis of acupuncture and moxibustion effect molecules,and provide ideas and methods for clinical diagnosis and treatment of diseases and evaluation of efficacy.

Objective Based on the phosphatidylinositol kinase(PI3K)/protein kinase B(AKT)pathway and M2 polarization of macrophages,the effects of polydatin on malignant phenotype of osteosarcoma cells were investigated.Methods The macrophages were divided into M0 group,M2 group,polydatin group,and polydatin+PI3K pathway agonist 740Y-P group.The expression of arginase-1(Arg-1)and CD206 mRNAs and proteins in macrophages were detected by RT-qPCR and Western blot.Western blot was used to detect p-PI3K/PI3K and p-AKT/AKT protein expression in macrophages.The osteosarcoma cells MG-63 were divided into control group,control group(RPMI-1640 medium culture),CM-M0 group(M0 group M0 macrophage supernatant is collected for osteosarcoma cells),CM-M2 group(M2 group M2 macrophage supernatant is collected for osteosarcoma cells),CM-polydatin group(M2 macrophage supernatant in the polydatin group was collected on osteosarcoma cells),and CM-polydatin+740Y-P group(M2 macrophage supernatant was collected on osteosarcoma cells in the CM-polydatin+740Y-P group).Edu staining assay was used to detect the proliferation of MG-63 cells.Scratches and Transwell assays were used to detect the migration and invasion of MG-63 cells.Results Compared with the M0 group,the Arg-1,CD206,IL-10,p-PI3K/PI3K,p-AKT/AKT increased in the M2 group significantly(P<0.05).Compared with the M2 group,the Arg-1,CD206,IL-10,p-PI3K/PI3K,p-AKT/AKT decreased in the polydatin group significantly(P<0.05).Compared with the polydatin group,the Arg-1,CD206,IL-10,p-PI3K/PI3K,p-AKT/AKT increased in the polydatin+740Y-P group significantly(P<0.05).Compared with the CM-M0 group,the proportion of Edu-positive cells in the CM-M2 group increased significantly(P<0.05),and MG-63 cell migration and invasion rates increased significantly(P<0.05).Compared with the CM-M2 group,the proportion of Edu-positive cells in the CM-polydatin group decreased significantly(P<0.05),and MG-63 cell migration and invasion rates decreased significantly(P<0.05).Compared with the CM-polydatin group,the proportion of Edu-positive cells in the CM-polydatin+740Y-P group increased significantly(P<0.05),and MG-63 cell migration and invasion rates increased significantly(P<0.05).Conclusion Polygonum cuspidatum can inhibit the proliferation,migration and invasion of osteosarcoma cells by inhibiting the M2 polarization of macrophages,and its mechanism may be related to the inhibition of PI3K/AKT signaling pathway.

Objective To investigate the effect of resveratrol(RES)on the function of osteoarthritis synovioblast(OA-FLS)and its molecular mechanism.Methods The purchased primary synovial cells were cultured to the third generation,and OA-FLS cell model was induced by IL-1β.The activity of FLS cells was observed by CCK-8 assay,and the appropriate RES intervention concentration was selected.The cells were divided into Control group,OA-FLS group,Low-RES group(1μmol/L)and High-RES group(5μmol/L).The effects of RES on the migration and invasion of OA-FLS were detected by cell scratch assay and Transwell assay.The effect of RES on OA-FLS apoptosis was detected by flow cytometry.The effects of RES on the expression of inflammatory factors in OA-FLS and re-lated proteins in NF-κB signaling pathway were detected by Western blot.Results The results of CCK-8 experiment showed that the concentration of RES above 10μmol/L significantly inhibited the viability of FLS cells,and the concentration of RES above 1 μmol/L and 5μmol/L had no significant effect on the viability of FLS cells.Compared with the Control group,the migration and invasion ability of OA-FLS group was enhanced,while the apoptosis ability was weakened,and the migration,invasion and apoptosis ability of OA-FLS group was significantly reversed after RES intervention.Western blot results showed that,compared with the Control group,the levels of inflammatory cytokines in OA-FLS group were increased,and the NF-κB signaling pathway was activated,while the treatment of OA-FLS with different concentrations of RES could reduce the expression of inflammatory cytokines and inhibit the transcription of nuclear fac-tors.Conclusion RES inhibited migration and invasion of OA-FLS,promoted apoptosis,and decreased the expression of inflammatory factors of OA-FLS,which may be related to the inhibition of NF-κB signaling pathway by RES.

Objective:To establish a method of dual nueleic acid test strips for rapid detection of Bacillus cereus cytK and nhe toxin genes based on polymerase chain reaction(PCR)and colloidal gold technique,and to evaluate its specificity,sensitivity,repeatability and stability.Methods:Bacillus cereus DNA was extracted by boiling method.Specific primers were designed with Bacillus cereus cytK and nhe as the target genes.Clonal transformation was used to identify the PCR products.The optimal labeling amounts of colloidal gold-labeled streptavidin,quality control line(C line),cytK detection line(T1)and nhe detection line(T2)were determined.The nucleic acid test strips were assembled and its specificity,sensitivity,reproducibility and stability were evaluated.Results:The DNA concentration of Bacillus cereus was 248 mg·L-1,and the purities were 1.8-2.0.After cloning and plasmid sequencing,the similarities between the PCR products and the sequences of cytK and nhe registered in the GenBank database were 100％.Under the condition of pH 7.0,the optimal amount of streptavidin labeling per 200 μL of colloidal gold solution was 6.0 μL;the optimal marking amount was 2.00 g·L-1 for the quality control line(C line),0.550 g·L-1 for cytK gene detection line(T1)and 0.2 g·L-1 for nhe gene detection line(T2).In the specificity test,positive result on the test strips was seen only for Bacillus cereus,and no cross-reactivity was observed for Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa and Bacillus subtilis,which were consistent with the electrophoresis results.Sensitivity assay showed that even when DNA concentration was reduced to 10-2 mg·L-1,three bands(C line,T1 line and T2 line)could be observed,and the detection limit of the test strip was one-tenth of agarose gel electrophoresis(10-1 mg·L-1).The nucleic acid test strips were verified by different operators in different laboratories,and the results were consistent.The stability of the test strips was verified at the 6th,9th and 12th months,and the results showed good stability.Conclusion:The dual nucleic acid test strip method established in this study can simutaneously detect the cytK and nhe toxin genes of Bacillus cereus with high sensitivity and specificity,achieving short-term visual detection.

Objective:To establish a rapid detection method for pathogenic microorganisms by combining loop-mediated isothermal amplification(LAMP)and clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 12a(Cas12a)(CRISPR-Cas12a)system,and to evaluate its efficacy for detecting the thermolabile hemolysin(tlh)gene of Vibrio parahaemolyticus(Vp).Methods:Using the tlh gene of Vp as the target gene,LAMP primers and CRISPR RNA(crRNA)were designed to construct and optimize the optimal concentration ratio of each component in the LAMP-CRISPR detection system.Bacillus cereus,Staphylococcus aureus,and Escherichia coli were used as control groups,and the specificity,sensitivity,reproducibility and positive conformity rate were verified to establish a rapid LAMP-CRISPR/Cas12a method for detecting the tlh gene of Vp.Results:The method specifically detected Vp,while Bacillus cereus,Staphylococcus aureus,and Escherichia coli yielded negative results.The DNA extraction concentration of Vp was 190.67 mg·L-1 with an A(260)/(A280)ratio of 1.84.Under the reaction conditions of 37℃ with 80 cycles for 40 min using quantitative PCR(qPCR)method,when the concentrations of Cas12a protein and crRNA in the LAMP-CRISPR/Cas12a system were 50 nmol·L-1,the visual brightness and relative fluorescence intensity peaks were high.The sensitivity of LAMP CRISPR/Cas12a for detecting Vp DNA concentration could reach 10-6 mg·L-1.The reproducibility test results showed that different experimenters had consistent results in different experimental environments and times.Conclusion:The established LAMP-CRISPR/Cas12a method can rapidly detect the tlh gene of Vp with high sensitivity and specificity,and can achieve short-term visual detection in the field.

Objective: To improve the efficiency and standardization of preoperative anemia diagnosis and treatment by establishing a systematic intelligent management platform for preoperative anemia. Methods: A multidisciplinary collaborative model was adopted to develop a preoperative anemia management system that integrates intelligent early warning, standardized treatment pathways, and quality control. The system utilizes natural language processing technology to automatically capture laboratory data and establish evidence-based medical decision support functions. A pre-post study design was employed to compare changes in preoperative anemia screening rates, preoperative anemia intervention rates, reasonable use of iron supplements, and perioperative red blood cell transfusion rates before and after system implementation. Results: After system implementation, the standardization of anemia diagnosis and treatment significantly improved: 1) Screening effectiveness: The anemia screening rate increased to 50.00% (an increase of 27.24%); 2) Intervention effectiveness: The anemia treatment rate rose to 56.30% (an increase of 14.02%); 3) Treatment standardization: The reasonable use rate of iron supplements increased to 55.33% (an increase of 21.02%); the red blood cell transfusion rate decreased to 18.29% (a decrease of 4.07%), and the amount of red blood cell transfusions was reduced by 291 units. Conclusion: This system achieves full-process management of preoperative anemia through information technology, significantly enhancing the standardization of diagnosis and treatment as well as intervention effectiveness, providing an effective solution for perioperative anemia management.