BACKGROUND:Mitochondrial calcium uptake 1 (MICU1) is one of the important molecules to maintain the mitochondrial calcium homeostasis. The regulation of MICU1 to mitochondrial calcium homeostasis may play an important role in diabetic cardiomyopathy, but the underlying mechanism remains unclear.
OBJECTIVE:To construct a lentiviral vector carrying MICU1 gene to transfect H9C2 cel s, and then to assess MICU1 level in H9C2 cel s thereby establishing a platform for researching the occurrence and development of diabetic cardiomyopathy at a cel ular level.
METHODS:DNA fragments of MICU1 were amplified by PCR, cleaved with Spe I, EcoR I and cloned into the lentiviral vector pRRLsin.CMV.eGFP to construct pRRLsin.CMV.MICU1-eGFP vector. 293T cel s were co-transfected with recombined pCMVDR8.91 and pCMV-VSVG to produce pRRLsin.CMV.MICU1-eGFP lentiviral viruses, and then used to infect H9C2 cel s. mRNA and protein expressions of MICU1 in the transfected H9C2 cel s were evaluated by real-time PCR and western blot assay. Mitochondrial calcium level in Rhod-2-stained H9C2 cel s was tested under confocal microscope.
RESULTS AND CONCLUSION:The recombinant inducible lentiviral vector containing MICU1 gene was successful y constructed. 293T could express green fluorescent protein with increased MICU1 level after pRRLsin.CMV.MICU1-eGFP transfection. The mRNA and protein expressions of MICU1 in the infected H9C2 group were obviously up-regulated compared with the other groups. MICU1 could remarkably improve the mitochondrial calcium level under Rhod-2 staining. These results show that pRRLsin.CMV.MICU1-eGFP lentiviral viruses are efficient to transfect H9C2 cel s, which wil be powered to lay a foundation for the immortalized cel line establishment.

To study the effect of essential MCU regulator (EMRE) on myocardial ischemia injury in experimental mice with underlining mechanism. Methods: Myocardial EMRE expression was up-regulated by EMRE adenovirus (Ad-EMRE) injection in mice myocardium tissue. Our research included in 3 groups: Sham operation group, sham mice received myocardium injection of eGFP adenovirus (Ad-eGFP); Myocardial infarction (MI) control group, the mice received Ad-eGFP injection and 48 hours later had coronary LAD ligation to establish MI model; MI treatment group, MI mice received Ad-EMRE injection. All animals were treated in 3 weeks. Mice cardiac function was examined by ultrasound; cardiomyocyte hypertrophy was evaluated by wheat germ agglutinin (WGA) staining, collagen fibrosis was measured by Masson staining, cell apoptosis was determined by TUNEL assay, protein expressions of EMRE, caspase-3 and caspase-9 were detected by Western blot analysis and mitochondrial reactive oxygen species (ROS) was assayed by MitoSOX fluorescence probe. Results: Compared with MI control group, MI treatment group showed the worse cardiac function, aggravated cardiac hypertrophy and elevated collagen fibrosis; in addition, MI treatment group had obviously increased cardiomyocyte apoptosis and increased protein expressions of Caspase-3, Caspase-9 and more mitochondrial ROS production. Conclusion: Over expressed EMRE can increase mitochondrial ROS production, induce cardiomyocyte apoptosis and therefore, aggravate myocardial ischemia injury in experimental mice.

Objective: Explore the method for volumetric measurement of alveolar bone defect. Methods: This study applied 2 advanced preoperative volume measurement methods: three-dimensional (3D) printing and computer-aided engineering (CAE). Twenty-six unilateral alveolar cleft patients were enrolled in this study from April 2015 to December 2016. Their computed tomographic data were sent to 3D printing and CAE software. A simulated graft was used on the 3D-printed model, and the graft volume was measured by water displacement. The volume calculated by CAE software used mirror-reverses technique. Results: The volume of alveolar bone defect could be detected by both methods. The average volume of the simulated bone grafts by 3D-printed models was 1.61 ml, a little higher than the mean volume of 1.60 ml calculated by CAE software. The difference between the 2 volumes was from -0.34 ml to 0.54 ml. The paired Student t test showed no statistically significant difference between the volumes derived from the 2 methods. Conclusions This study demonstrated that the volume of alveolar bone defect is about 1.6ml in unilateral alveolar cleft patients aged 9-12 years. The mirror-reversed technique by CAE software is as accurate as the simulated operation on 3D-printed models. These findings further validate the use of 3D printing and CAE technique in alveolar defect repairing.

Objective:To construct a whole course management and follow-up platform, named Care Plan, for patients undergoing breast reconstruction surgery, and to evaluate its initial clinical applications.Methods:From June 2023 to January 2024, patients who underwent breast reconstruction surgery at Peking Union Medical College Hospital were selected as the study subjects and enrolled in the Care Plan platform for whole course management. The platform comprised five modules: patient education, self-assessment, medical assessment, medical follow-up, and doctor-patient dialogue, each with corresponding sub-modules and specific components. Each patient was assigned to a case manager who provided one-on-one assessment, sent patient education materials, and conducted follow-ups. The preliminary application of the Care Plan platform was evaluated based on the completeness of patient data, patient education material reading status, doctor-patient dialogue topics, and follow-up responses. At 3 months postoperatively, patients were followed up and further divided into patient education group(reading at least an article on patient educational materials) and control group based on their reading status of educational materials. Patients rated their satisfaction with the surgery using a 5-point questionnaire, including five indicators: expected breast reconstruction outcome, expected surgical type, completeness of information acquisition, breast symmetry, and breast softness. Statistical analysis was performed using SPSS 27.0.1 software, with count data expressed as frequencies and (or) percentages, normally distributed measurement data expressed as Mean±SD, intergroup comparisons conducted using t-test, and P<0.05 indicating statistical significance. Results:A total of 147 female patients aged 16 to 63 years with a body mass index of 16.6 to 31.3 kg/m 2 were ultimately included. All 147 patients completed the content of the self-assessment and medical assessment modules, with relatively complete data. Of the 133 patients who received patient education materials sent through the platform, 38 (28.6%) read the materials, with the sub-module on the introduction and choices of the reconstruction surgery (37.3%, 94/252 ) being the most frequently viewed. A total of 24 patients (16.3%, 24/147 ) raised 45 questions using the doctor-patient dialogue module, with the highest proportion related to consultation issues and complications, each accounting for 22.2% (10/45). Twenty-three patients completed the 3-month follow-up, with 12 in the patient education group and 11 in the control group. The patient education group scored higher than the control group in terms of completeness of information acquisition [(4.58±0.51) points vs. (3.91±0.70) points, t=-2.65, P=0.015], while scoring lower in breast symmetry [(3.92±0.79) points vs. (4.63±0.67) points, t=2.33, P=0.030]. There was no significant difference between the two groups in terms of expected breast reconstruction outcome, expected surgical type, and breast softness scores ( P>0.05 for all). Conclusion:The Care Plan platform for whole course management and follow-up of breast reconstruction can assist medical staff in formulating patient-centered management plans, improve patients’ understanding of surgical information, and aid patients in making decisions regarding breast reconstruction methods.

Objective:To construct a whole course management and follow-up platform, named Care Plan, for patients undergoing breast reconstruction surgery, and to evaluate its initial clinical applications.Methods:From June 2023 to January 2024, patients who underwent breast reconstruction surgery at Peking Union Medical College Hospital were selected as the study subjects and enrolled in the Care Plan platform for whole course management. The platform comprised five modules: patient education, self-assessment, medical assessment, medical follow-up, and doctor-patient dialogue, each with corresponding sub-modules and specific components. Each patient was assigned to a case manager who provided one-on-one assessment, sent patient education materials, and conducted follow-ups. The preliminary application of the Care Plan platform was evaluated based on the completeness of patient data, patient education material reading status, doctor-patient dialogue topics, and follow-up responses. At 3 months postoperatively, patients were followed up and further divided into patient education group(reading at least an article on patient educational materials) and control group based on their reading status of educational materials. Patients rated their satisfaction with the surgery using a 5-point questionnaire, including five indicators: expected breast reconstruction outcome, expected surgical type, completeness of information acquisition, breast symmetry, and breast softness. Statistical analysis was performed using SPSS 27.0.1 software, with count data expressed as frequencies and (or) percentages, normally distributed measurement data expressed as Mean±SD, intergroup comparisons conducted using t-test, and P<0.05 indicating statistical significance. Results:A total of 147 female patients aged 16 to 63 years with a body mass index of 16.6 to 31.3 kg/m 2 were ultimately included. All 147 patients completed the content of the self-assessment and medical assessment modules, with relatively complete data. Of the 133 patients who received patient education materials sent through the platform, 38 (28.6%) read the materials, with the sub-module on the introduction and choices of the reconstruction surgery (37.3%, 94/252 ) being the most frequently viewed. A total of 24 patients (16.3%, 24/147 ) raised 45 questions using the doctor-patient dialogue module, with the highest proportion related to consultation issues and complications, each accounting for 22.2% (10/45). Twenty-three patients completed the 3-month follow-up, with 12 in the patient education group and 11 in the control group. The patient education group scored higher than the control group in terms of completeness of information acquisition [(4.58±0.51) points vs. (3.91±0.70) points, t=-2.65, P=0.015], while scoring lower in breast symmetry [(3.92±0.79) points vs. (4.63±0.67) points, t=2.33, P=0.030]. There was no significant difference between the two groups in terms of expected breast reconstruction outcome, expected surgical type, and breast softness scores ( P>0.05 for all). Conclusion:The Care Plan platform for whole course management and follow-up of breast reconstruction can assist medical staff in formulating patient-centered management plans, improve patients’ understanding of surgical information, and aid patients in making decisions regarding breast reconstruction methods.