In response to issues in diagnostics experiment teaching for intelligent medical engineering in medical colleges and universities, such as the disconnection between theory and practice, the teaching content emphasizing "medicine" rather than "engineering", the lack of interaction between teachers and students, and the single teaching evaluation method, this study introduced the outcome-based education concept. According to the graduation requirements supported by the course, the experiment teaching reform ideas were adjusted from three aspects: setting curriculum objectives based on learning outcomes, reforming teaching strategies to achieve learning outcomes, and assessing learning outcomes to promote continuous improvement. The results of the reform show that students' satisfaction with the content, methods, and assessment of experiment teaching all exceeded 86.66%. The reform successfully integrated medicine and engineering, virtual and practical training, and teaching and ideological and political education. The reform also significantly improved the teaching effectiveness and talent cultivation level of experimental courses from the three dimensions of knowledge learning, ability improvement, and value shaping.

Objective:To investigate the effect and underlying mechanism of sonodynamic therapy (SDT) on inflammation-related atrial fibrillation (AF) susceptibility.Methods:Lipopolysaccharide (LPS)-stimulated mouse and HL-1 mouse atrial myocyte models were used. (1) In vivo study: experimental groups included control, LPS, LPS+SDT, and SDT groups, with 20 mice in each group. Atrial fibrillation inducibility and duration were assessed by electrical stimulation. Western blot was used to analyze atrial expression of NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), interleukin (IL)-1β, and IL-18. Immunohistochemistry was used to detect calcium voltage-gated channel subunit alpha1 C (CACNA1C) expression. (2) In vitro study: cell counting kit-8 (CCK-8) and Western blot were used to determine the optimal and safe LPS concentration. The safe incubation condition for the sonosensitizer sinoporphyrin sodium was determined by CCK-8 and fluorometry. An LPS-induced inflammatory model in HL-1 atrial myocytes was used, with experimental groups including control, LPS, LPS+SDT, LPS+sinoporphyrin sodium, and LPS+ultrasound groups. NLRP3 was overexpressed using plasmid transfection, with experimental groups including control, NLRP3 plasmid, negative control plasmid, and NLRP3 plasmid+SDT groups. SDT was applied to LPS-stimulated or NLRP3-overexpressing HL-1 cells. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to measure mRNA and protein levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Cleaved Caspase-1, IL-1β, IL-18, and CACNA1C. The NLRP3 inhibitor MCC950 was used to validate the relationship of NLRP3 and CACNA1C. The experimental groups included control, LPS, LPS+MCC950, and MCC950 groups. Intracellular reactive oxygen species (ROS) levels were detected using the probe DCFH-DA, and the ROS scavenger N-acetyl-L-cysteine (NAC) was used to test if the effects of SDT was ROS-dependent.Results:(1) In vivo: The LPS+SDT group exhibited a lower incidence of atrial fibrillation induction and a shorter duration of atrial fibrillation compared to the LPS group(both P<0.05). Protein expression levels of NLRP3 and IL-1β were lower than those in the LPS group (all P<0.05), while the expression of CACNA1C subunit tended to increase relative to the LPS group ( P>0.05). (2) In vitro: The safe concentration of LPS for administration was ≤20 μg/ml, with an optimal pro-inflammatory concentration of 4 μg/ml. The safe concentration of sinoporphyrin sodium for administration was 0.4 μmol/L, with an optimal incubation time of 4 hours. Compared to the LPS group or NLRP3 plasmid group, the LPS+SDT group or NLRP3 plasmid+SDT group exhibited lower expression levels of NLRP3, ASC, Cleaved Caspase-1, IL-1β, and IL-18, and higher mRNA and protein levels of CACNA1C (all P<0.05). The LPS+MCC950 group had higher CACNA1C protein expression than the LPS group ( P<0.05). SDT increased intracellular ROS levels, and NAC blocked the regulatory effects of SDT on NLRP3 and CACNA1C. Conclusion:SDT reduces atrial fibrillation susceptibility in mice by inhibiting NLRP3 inflammasome activation in atrial cardiomyocytes, thereby upregulating the L-type calcium channel subunit CACNA1C.

After hepatitis B virus (HBV) infection, numerous patients will develop serious sequelae such as liver failure and liver cancer. Finding a way to eradicate HBV has emerged as a top priority for the treatment of HBV infection. Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system represents a potent tool for genome manipulation. Many studies have confirmed the capability of the CRISPR/Cas9 gene editing system to remove the covalently closed circular DNA of HBV in cells, which has great potential for application in the treatment of chronic HBV infection. This article summarizes the application status and research progress of CRISPR/Cas9 system in anti-HBV infection, and explores the challenges that may be encountered.

Objective:To investigate the effect and underlying mechanism of sonodynamic therapy (SDT) on inflammation-related atrial fibrillation (AF) susceptibility.Methods:Lipopolysaccharide (LPS)-stimulated mouse and HL-1 mouse atrial myocyte models were used. (1) In vivo study: experimental groups included control, LPS, LPS+SDT, and SDT groups, with 20 mice in each group. Atrial fibrillation inducibility and duration were assessed by electrical stimulation. Western blot was used to analyze atrial expression of NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), interleukin (IL)-1β, and IL-18. Immunohistochemistry was used to detect calcium voltage-gated channel subunit alpha1 C (CACNA1C) expression. (2) In vitro study: cell counting kit-8 (CCK-8) and Western blot were used to determine the optimal and safe LPS concentration. The safe incubation condition for the sonosensitizer sinoporphyrin sodium was determined by CCK-8 and fluorometry. An LPS-induced inflammatory model in HL-1 atrial myocytes was used, with experimental groups including control, LPS, LPS+SDT, LPS+sinoporphyrin sodium, and LPS+ultrasound groups. NLRP3 was overexpressed using plasmid transfection, with experimental groups including control, NLRP3 plasmid, negative control plasmid, and NLRP3 plasmid+SDT groups. SDT was applied to LPS-stimulated or NLRP3-overexpressing HL-1 cells. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to measure mRNA and protein levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Cleaved Caspase-1, IL-1β, IL-18, and CACNA1C. The NLRP3 inhibitor MCC950 was used to validate the relationship of NLRP3 and CACNA1C. The experimental groups included control, LPS, LPS+MCC950, and MCC950 groups. Intracellular reactive oxygen species (ROS) levels were detected using the probe DCFH-DA, and the ROS scavenger N-acetyl-L-cysteine (NAC) was used to test if the effects of SDT was ROS-dependent.Results:(1) In vivo: The LPS+SDT group exhibited a lower incidence of atrial fibrillation induction and a shorter duration of atrial fibrillation compared to the LPS group(both P<0.05). Protein expression levels of NLRP3 and IL-1β were lower than those in the LPS group (all P<0.05), while the expression of CACNA1C subunit tended to increase relative to the LPS group ( P>0.05). (2) In vitro: The safe concentration of LPS for administration was ≤20 μg/ml, with an optimal pro-inflammatory concentration of 4 μg/ml. The safe concentration of sinoporphyrin sodium for administration was 0.4 μmol/L, with an optimal incubation time of 4 hours. Compared to the LPS group or NLRP3 plasmid group, the LPS+SDT group or NLRP3 plasmid+SDT group exhibited lower expression levels of NLRP3, ASC, Cleaved Caspase-1, IL-1β, and IL-18, and higher mRNA and protein levels of CACNA1C (all P<0.05). The LPS+MCC950 group had higher CACNA1C protein expression than the LPS group ( P<0.05). SDT increased intracellular ROS levels, and NAC blocked the regulatory effects of SDT on NLRP3 and CACNA1C. Conclusion:SDT reduces atrial fibrillation susceptibility in mice by inhibiting NLRP3 inflammasome activation in atrial cardiomyocytes, thereby upregulating the L-type calcium channel subunit CACNA1C.

In response to issues in diagnostics experiment teaching for intelligent medical engineering in medical colleges and universities, such as the disconnection between theory and practice, the teaching content emphasizing "medicine" rather than "engineering", the lack of interaction between teachers and students, and the single teaching evaluation method, this study introduced the outcome-based education concept. According to the graduation requirements supported by the course, the experiment teaching reform ideas were adjusted from three aspects: setting curriculum objectives based on learning outcomes, reforming teaching strategies to achieve learning outcomes, and assessing learning outcomes to promote continuous improvement. The results of the reform show that students' satisfaction with the content, methods, and assessment of experiment teaching all exceeded 86.66%. The reform successfully integrated medicine and engineering, virtual and practical training, and teaching and ideological and political education. The reform also significantly improved the teaching effectiveness and talent cultivation level of experimental courses from the three dimensions of knowledge learning, ability improvement, and value shaping.

After hepatitis B virus (HBV) infection, numerous patients will develop serious sequelae such as liver failure and liver cancer. Finding a way to eradicate HBV has emerged as a top priority for the treatment of HBV infection. Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system represents a potent tool for genome manipulation. Many studies have confirmed the capability of the CRISPR/Cas9 gene editing system to remove the covalently closed circular DNA of HBV in cells, which has great potential for application in the treatment of chronic HBV infection. This article summarizes the application status and research progress of CRISPR/Cas9 system in anti-HBV infection, and explores the challenges that may be encountered.

Organ preservation fluid could mitigate cold ischemia injury and maintain normal function of the grafts. At present, how to reduce a series of injury caused by cold ischemia of donor liver and improve the preservation quality of grafts are the hot and challenging spots in this field. Currently, preservation fluid in clinical practice has not achieved ideal preservation effect, especially for the protection of marginal donor organs. In the context of severe donor shortage, the key solution is still to explore the optimal preservation protocol for donor liver to prevent grafts from cold ischemia injury. In this article, the mechanism of donor liver injury during cold ischemia, the classification and evolution of donor liver preservation fluid were summarized, the development direction and challenges of donor liver preservation fluid were discussed, aiming to provide novel ideas and references for the research and development of donor liver preservation fluid.

Objective To investigate the safety and efficacy of different surgical sequences in the ultrasound-guided endovenous microwave ablation combined with foam sclerotherapy for the treatment of primary great saphenous varicose veins.Method A total of 80 patients with great saphenous varicose vein admitted in the affiliated hospital of Jiangsu University,from January 2022 to January 2023 were selected.Patients were divided into observation group and control group according to different operation order,with 40 cases in each group.The control group was treated with ultrasound-guided micro wave ablation of the main saphenous vein was performed first,followed by superficial calf vein foam sclerotherapy injection and local small incision point extraction,and the observation group was treated with superficial calf vein foam sclerotherapy injection and local small incision point extraction first,followed by ultrasound-guided microwave ablation of the main saphenous vein was performed.Perioperative relevant indicators at the 1st week of the two groups were counted,and the incidence of hematoma,ecchymosis,induration,skin burn,thrombotic superficial phlebitis,and endovenous heat induced thrombosis at the 1st week after surgery.The venous clinical severity score and chronic venous insufficiency quality of life at the 3rd and 6th month after surgery were compared between the two groups.VCSS and CIVIQ were used to evaluate the postoperative recovery of patients with varicose veins.Six months after the operation,the recurrence rate of great saphenous vein was compared by color Doppler ultrasonography.Result The operation time of the two groups was(68.13±3.34)min and(66.83±3.19)min,respectively.The intraoperative blood loss was(15.35±2.63)ml and(14.83±2.66)ml,respectively.The underground activity time was(14.35±3.34)hours and(13.60±2.63)hours,respectively.The length of hospitalization was(2.93±0.52)days and(3.15±0.61)days,respectively.There was statistical significance between the two groups(P＜0.05).The preoperative VCSS of the two groups were 4.08±1.37 and 4.23±1.33,respectively,3 months after surgery were 3.00±0.59 and 3.03±0.61,respectively,and 6 months after surgery were 2.20±1.17 and 2.35±0.96,respectively.The preoperative CIVIQ of the two groups were 79.63±5.41 and 80.03±7.44,respectively,3 months after operation was 69.90±2.98 and 70.43±3.55,respectively,the 6-month CIVIQ was 59.05±3.79 and 58.00±4.66,respectively.There was no statistical significance between the two groups(P＞0.05).The incidence of adverse events[hematoma(0 vs 0),ecchymosis(12.5％vs 15.0％),sclerosis(10.0％vs 7.5％),skin burns(0 vs 0),thrombosed superficial phlebitis(12.5％vs 17.5％),and thermal ablation-induced thrombosis(10.0％vs 5.0％)]in the patients of the two groups in the 1-week period after the procedure were compared,and the difference were statistically non-significant(P＞0.05).Comparison of trunk recanalisation rate(5.0％vs 2.5％)at 6 months after surgery,the difference was not statistically significant(P＞0.05).Conclusion There is no significant difference in the efficacy of the two procedures in the treatment of primary saphenous varicose veins,with a high degree of safety,both of which are worthy of clinical promotion.

Objective To optimize the culture and fermentation conditions of the Penicillium aurantiocandidum Z12 strain, a fungal strain with molluscicidal actions against Oncomelania hupensis, so as to provide the basis for the research and development of molluscicidal active substances from the P. aurantiocandidum Z12 strain and its fermentation broth and large-scale fermentation. Methods The carbon source, nitrogen source and mineral salts were identified in the optimal culture medium for the P. aurantiocandidum Z12 strain with a single-factor experiment to determine the best fermentation condition for the P. aurantiocandidum Z12 strain. Factors that significantly affected the growth of the P. aurantiocandidum Z12 strain were identified using the Plackett-Burman design, and the best range of each factor was determined using the steepest climb test. Response surface analyses of temperature, pH value, seeding amount and liquid-filling quantity were performed using the Box-Behnken design to create a regression model for fermentation of the P. aurantiocandidum Z12 strain to identify the optimal culture medium. Results Single-factor experiment preliminarily identified the best culture medium and conditions for the P. aurantiocandidum Z12 strain as follows: sucrose as the carbon source at approximately 20 g/L, tryptone as the nitrogen source at approximately 5 g/L, K₂HPO₄ as the mineral salt at approximately 5 g/L, initial pH at approximately 8, temperature at approximately 28 °C, seeding amount at approximately 6%, and liquid-filling quantity at approximately 50 mL/100 mL. Plackett-Burman design showed that factors that significantly affected the growth of the P. aurantiocandidum Z12 strain included temperature (t = −5.28, P < 0.05), seeding amount (t = 5.22, P < 0.05), pH (t = −4.30, P < 0.05) and liquid-filling quantity (t = −4.39, P < 0.05). Steepest climb test showed the highest mycelial growth at pH of 7.5, seeding amount of 8%, and liquid-filling quantity of 40 mL/100 mL, and this condition was selected as the central point of response surface analysis for the subsequent optimization of fermentation conditions. Response surface analyses using the Box-Behnken design showed that the optimal conditions for fermentation of the P. aurantiocandidum Z12 strain included sucrose at 15 g/L, tryptone at 5 g/L, K₂HPO₄ at 5 g/L, temperature at 28.2 °C, pH at 7.5, seeding amount at 10%, and liquid-filling quantity at 35.8 mL/100.0 mL, resulting in 0.132 g yield of the P. aurantiocandidum Z12 strain. Conclusion The optimal culture condition for the P. aurantiocandidum Z12 strain has been identified, and the optimized culture medium and fermentation condition may effectively improve the fermentation yield of the P. aurantiocandidum Z12 strain.

New drug discovery is under growing pressure to satisfy the demand from a wide range of domains, especially from the pharmaceutical industry and healthcare services. Assessment of drug efficacy and safety prior to human clinical trials is a crucial part of drug development, which deserves greater emphasis to reduce the cost and time in drug discovery. Recent advances in microfabrication and tissue engineering have given rise to organ-on-a-chip, an in vitro model capable of recapitulating human organ functions in vivo and providing insight into disease pathophysiology, which offers a potential alternative to animal models for more efficient pre-clinical screening of drug candidates. In this review, we first give a snapshot of general considerations for organ-on-a-chip device design. Then, we comprehensively review the recent advances in organ-on-a-chip for drug screening. Finally, we summarize some key challenges of the progress in this field and discuss future prospects of organ-on-a-chip development. Overall, this review highlights the new avenue that organ-on-a-chip opens for drug development, therapeutic innovation, and precision medicine.