Objective:To evaluate the relationship between nuclear receptor subfamily 1 group D member 1 (Rev-erbα) and ferroptosis in cardiomyocytes subjected to high-fat/high-glucose (HFHG) and hypoxia-reoxygenation (H/R) injury.Methods:H9c2 cardiomyocytes were cultured under normal conditions. The cells were divided into 4 groups ( n=13 each) using a random number table method: control group (C group), H/R group, HFHG group and HFHG+ H/R1 group. The cells were divided into 3 groups ( n=17 each) using a random number table method: HFHG+ H/R2 group, negative control siRNA + HFHG + H/R group (si-NC+ HFHG+ H/R group), and Rev-erbα gene knockdown + HFHG + H/R group (si-Rev-erbα+ HFHG+ H/R group). The cardiomyocyte model of HFHG combined with H/R injury was established by incubating cells with HFHG medium for 12 h, followed by 6 h of hypoxia and 2 h of reoxygenation. Rev-erbα gene was knocked down using siRNA technology. Cell viability was assessed using CCK-8 and Calcein AM/PI live-dead cell double staining kits. The expression of Rev-erbα, acyl-CoA synthetase long-chain family member 4 (ACSL4), and nuclear receptor coactivator 4 (NCOA4) was detected by Western blot. The levels of lipid peroxide (LPO) were measured by flow cytometry. Results:Compared with C group, the cell viability was significantly decreased, and the expression of Rev-erbα, ACSL4 and NCOA4 was up-regulated in HFHG, H/R and HFHG+ H/R1 groups( P<0.05). Compared with HFHG group or H/R group, the cell viability was significantly decreased, and the expression of Rev-erbα, ACSL4 and NCOA4 was up-regulated in HFHG+ H/R1 group ( P<0.05).There were no significant differences in the cell viability, levels of LPO, or expression of Rev-erbα, ACSL4 and NCOA4 between HFHG+ H/R2 group and si-NC+ HFHG+ H/R group ( P>0.05). Compared with HFHG+ H/R2 group, the cell viability was significantly increased, the levels of LPO were decreased, and the expression of Rev-erbα, ACSL4 and NCOA4 was down-regulated in si-Rev-erbα+ HFHG+ H/R group ( P<0.05). Conclusions:Rev-erbα participates in the process of HFHG and H/R injury to cardiomyocytes by negatively regulating ferroptosis.

Diterpenoid ferruginol is a key intermediate in biosynthesis of active ingredients such as tanshinone and carnosic acid.However, the traditional process of obtaining ferruginol from plants is often cumbersome and inefficient. In recent years, the increasingly developing gene editing technology has been gradually applied to the heterologous production of natural products, but the production of ferruginol in microbe is still very low, which has become an obstacle to the efficient biosynthesis of downstream chemicals, such as tanshinone. In this study, miltiradiene was produced by integrating the shortened diterpene synthase fusion protein,and the key genes in the MVA pathway were overexpressed to improve the yield of miltiradiene. Under the shake flask fermentation condition, the yield of miltiradiene reached about(113. 12±17. 4)mg·L~(-1). Subsequently, this study integrated the ferruginol synthase Sm CYP76AH1 and Sm CPR1 to reconstruct the ferruginol pathway and thereby realized the heterologous synthesis of ferruginol in Saccharomyces cerevisiae. The study selected the best ferruginol synthase(Il CYP76AH46) from different plants and optimized the expression of pathway genes through redox partner engineering to increase the yield of ferruginol. By increasing the copy number of diterpene synthase, CYP450, and CPR, the yield of ferruginol reached(370. 39± 21. 65) mg·L~(-1) in the shake flask, which was increased by 21. 57-fold compared with that when the initial ferruginol strain JMLT05 was used. Finally, 1 083. 51 mg·L~(-1) ferruginol was obtained by fed-batch fermentation, which is the highest yield of ferruginol from biosynthesis so far. This study provides not only research ideas for other metabolic engineering but also a platform for the construction of cell factories for downstream products.
Saccharomyces cerevisiae/genetics* ; Diterpenes/metabolism* ; Metabolic Engineering ; Fermentation ; Abietanes

Testicular histology based on testicular biopsy is an important factor for determining appropriate testicular sperm extraction surgery and predicting sperm retrieval outcomes in patients with azoospermia. Therefore, we developed a deep learning (DL) model to establish the associations between testicular grayscale ultrasound images and testicular histology. We retrospectively included two-dimensional testicular grayscale ultrasound from patients with azoospermia (353 men with 4357 images between July 2017 and December 2021 in The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China) to develop a DL model. We obtained testicular histology during conventional testicular sperm extraction. Our DL model was trained based on ultrasound images or fusion data (ultrasound images fused with the corresponding testicular volume) to distinguish spermatozoa presence in pathology (SPP) and spermatozoa absence in pathology (SAP) and to classify maturation arrest (MA) and Sertoli cell-only syndrome (SCOS) in patients with SAP. Areas under the receiver operating characteristic curve (AUCs), accuracy, sensitivity, and specificity were used to analyze model performance. DL based on images achieved an AUC of 0.922 (95% confidence interval [CI]: 0.908-0.935), a sensitivity of 80.9%, a specificity of 84.6%, and an accuracy of 83.5% in predicting SPP (including normal spermatogenesis and hypospermatogenesis) and SAP (including MA and SCOS). In the identification of SCOS and MA, DL on fusion data yielded better diagnostic performance with an AUC of 0.979 (95% CI: 0.969-0.989), a sensitivity of 89.7%, a specificity of 97.1%, and an accuracy of 92.1%. Our study provides a noninvasive method to predict testicular histology for patients with azoospermia, which would avoid unnecessary testicular biopsy.
Humans ; Male ; Azoospermia/diagnostic imaging* ; Deep Learning ; Testis/pathology* ; Retrospective Studies ; Adult ; Ultrasonography/methods* ; Sperm Retrieval ; Sertoli Cell-Only Syndrome/diagnostic imaging*

Objective To analyze the risk factors of permanent hypoparathyroidism(pHPP)after total thyroidectomy in patients with thyroid cancer and establish a nomogram prediction model.Methods A total of 245 patients with thyroid cancer who received total thyroidectomy in our hospital were enrolled between January 2020 and January 2024.According to presence or absence of postoperative pHPP,patients were divided into the pHPP group and the non-pHPP group.The influencing factors of postoperative pHPP in patients with thyroid cancer were analyzed by univariate and multivariate Logistic regression analysis.The nomogram prediction model for postoperative pHPP in patients with thyroid cancer was constructed and varified,and efficiency of the model was evaluated.Results In 245 patients with thyroid cancer,the incidence of pHPP within 6 months after surgery was 10.20%(25/245).Univariate analysis showed that there were significant differences in tumor size,surgical method,central lymph node dissection,use of nano carbon tracer,envelope invasion,parathyroid excision by mistake,Hashimoto thyroiditis,serum calcium and parathyroid hormone at 1 d after surgery between the two groups(P＜0.05),but there were no significant differences in gender,age,smoking,drinking,extraglandular invasion,parathyroid autologous transplantation,preoperative vitamin D or serum phosphorus at 1 d after surgery between the two groups(P＞0.05).Multivariate analysis showed that maximum tumor diameter≥4 cm,routine and open total thyroidectomy,central lymph node dissection,no use of nano carbon tracer and parathyroid excision by mistake were all independent risk factors for postoperative pHPP in patients with thyroid cancer(P＜0.05).Results of nomogram prediction model showed that C-index was 0.921,the corrected curve was close to ideal curve,and AUC of nomogram model for predicting postoperative pHPP was 0.926(95%CI:0.871-0.981).Conclusion The nomogram prediction model constructed based on independent risk factors of postoperative pHPP has good predictive efficiency in patients with thyroid cancer.

Objective To investigate the influence of body mass index(BMI)on individualized analgesic effect,inflammatory factors and safety of sufentanil in elderly patients after proximal femoral nail anti-rotation(PFNA)surgery.Methods A total of 161 elderly patients who received PFNA surgery in Nanjing Drum Tower Hospital Group Suqian Hospital from January 2022 to December 2023 were selected as study subjects.Patients with BMI<18.5 kg/m2 were set as Group A,patients with BMI ranging from 18.5 kg/m2 to 23.9 kg/m2 were set as Group B,patients with BMI ranging from 24.0 kg/m2 to 35.0 kg/m2 were set as Group C.After operation,individualized analgesia was performed with an intravenous patient-controlled analgesia pump of 2 μg/kg sufentanil based on body weight,and the postoperative pain degree,inflammatory factors,analgesia condition and adverse reactions of patients in the three groups were compared.Results At 8 hours,12 hours,24 hours and 48 hours after surgery,the pain visual analogue scale(VAS)scores of patietns in group B and group C were significantly lower than those in group A(P<0.05).At 12 hours,24 hours and 48 hours after surgery,the pain VAS scores of patients in group C were significantly lower than those in group B(P<0.05).The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)1 day after surgery of patients in group B and group C were significantly lower than those in group A(P<0.05).The pressing times of analgesia pump and the duration of obvious pain within 48 hours after surgery of patients in group A were significantly more/longer than those in group B and group C(P<0.05),and the analgesic satisfaction score was significantly lower than those in group B and group C(P<0.05).The total incidence of analgesic adverse reactions in group C was significantly higher than those in group A and group B(P<0.05).Conclusion BMI may affect the individualized analgesic effect,inflammatory factors and safety of sufentanil in elderly patients undergoing PFNA surgery.Underweight patients may have insufficient analgesia,while overweight or obese patients may have excessive analgesia,which may affect the analgesic safety.

Objective To explore the effects of different CO2 pneumoperitoneum pressures during surgery on overweight or obese patients with laparoscopic panhysterectomy in Trendelenburg position.Methods A total of 88 overweight or obese patients who underwent laparoscopic panhysterectomy were selected and randomly divided into the low-pressure group and the high-pressure group according to random number table method,with 44 patients in each group.All patients used CO2 as the pneumoperitoneum medium during surgery,with 10 mmHg of CO2 pneumoperitoneum pressure in the low-pressure group and 15 mmHg of CO2 pneumoperitoneum pressure in the high-pressure group.The operation-related indexes,respiratory indicators,nasopharyngeal temperature and adverse reactions of patients were compared between the two groups.Results There was no significant difference in the surgical time,blood loss,intraoperative infusion volume,incidence of organ injury,and laparotomy rate between the two groups(P>0.05).The recovery time of anesthesia of patients in the low-pressure group was shorter than that in the high-pressure group(P<0.05).At 30 minutes after the establishment of pneumoperitoneum,the peak airway pressure(Ppeak)and plateau pressure(Pplat)of patients in the high-pressure group were significantly higher than those in the low-pressure group(P<0.05),while the dynamic lung compliance(Cdyn)of patients in the high-pressure group was significantly lower than that in the low-pressure group(P<0.05).At 60 minutes after the establishment of pneumoperitoneum and the end of pneumoperitoneum,the nasopharyngeal temperature of patients in the high-pressure group were significantly lower than those in the low-pressure group(P<0.05).The incidences of intraoperative hypothermia and shivering in the high-pressure group were significantly higher than those in the low-pressure group(P<0.05).Conclusion Compared with 15 mmHg of CO2 pneumoperitoneum pressure,10 mmHg of CO2 pneumoperitoneum pressure on overweight or obese patients with laparoscopic panhysterectomy in Trendelenburg position does not increase surgical difficulty,and the patients has lower airway pressure,better lung compliance,fewer adverse reactions,and faster recovery,which can also avoid intraoperative hypothermia.

Objective To analyze the risk factors of permanent hypoparathyroidism(pHPP)after total thyroidectomy in patients with thyroid cancer and establish a nomogram prediction model.Methods A total of 245 patients with thyroid cancer who received total thyroidectomy in our hospital were enrolled between January 2020 and January 2024.According to presence or absence of postoperative pHPP,patients were divided into the pHPP group and the non-pHPP group.The influencing factors of postoperative pHPP in patients with thyroid cancer were analyzed by univariate and multivariate Logistic regression analysis.The nomogram prediction model for postoperative pHPP in patients with thyroid cancer was constructed and varified,and efficiency of the model was evaluated.Results In 245 patients with thyroid cancer,the incidence of pHPP within 6 months after surgery was 10.20%(25/245).Univariate analysis showed that there were significant differences in tumor size,surgical method,central lymph node dissection,use of nano carbon tracer,envelope invasion,parathyroid excision by mistake,Hashimoto thyroiditis,serum calcium and parathyroid hormone at 1 d after surgery between the two groups(P＜0.05),but there were no significant differences in gender,age,smoking,drinking,extraglandular invasion,parathyroid autologous transplantation,preoperative vitamin D or serum phosphorus at 1 d after surgery between the two groups(P＞0.05).Multivariate analysis showed that maximum tumor diameter≥4 cm,routine and open total thyroidectomy,central lymph node dissection,no use of nano carbon tracer and parathyroid excision by mistake were all independent risk factors for postoperative pHPP in patients with thyroid cancer(P＜0.05).Results of nomogram prediction model showed that C-index was 0.921,the corrected curve was close to ideal curve,and AUC of nomogram model for predicting postoperative pHPP was 0.926(95%CI:0.871-0.981).Conclusion The nomogram prediction model constructed based on independent risk factors of postoperative pHPP has good predictive efficiency in patients with thyroid cancer.

Objective:To evaluate the relationship between nuclear receptor subfamily 1 group D member 1 (Rev-erbα) and ferroptosis in cardiomyocytes subjected to high-fat/high-glucose (HFHG) and hypoxia-reoxygenation (H/R) injury.Methods:H9c2 cardiomyocytes were cultured under normal conditions. The cells were divided into 4 groups ( n=13 each) using a random number table method: control group (C group), H/R group, HFHG group and HFHG+ H/R1 group. The cells were divided into 3 groups ( n=17 each) using a random number table method: HFHG+ H/R2 group, negative control siRNA + HFHG + H/R group (si-NC+ HFHG+ H/R group), and Rev-erbα gene knockdown + HFHG + H/R group (si-Rev-erbα+ HFHG+ H/R group). The cardiomyocyte model of HFHG combined with H/R injury was established by incubating cells with HFHG medium for 12 h, followed by 6 h of hypoxia and 2 h of reoxygenation. Rev-erbα gene was knocked down using siRNA technology. Cell viability was assessed using CCK-8 and Calcein AM/PI live-dead cell double staining kits. The expression of Rev-erbα, acyl-CoA synthetase long-chain family member 4 (ACSL4), and nuclear receptor coactivator 4 (NCOA4) was detected by Western blot. The levels of lipid peroxide (LPO) were measured by flow cytometry. Results:Compared with C group, the cell viability was significantly decreased, and the expression of Rev-erbα, ACSL4 and NCOA4 was up-regulated in HFHG, H/R and HFHG+ H/R1 groups( P<0.05). Compared with HFHG group or H/R group, the cell viability was significantly decreased, and the expression of Rev-erbα, ACSL4 and NCOA4 was up-regulated in HFHG+ H/R1 group ( P<0.05).There were no significant differences in the cell viability, levels of LPO, or expression of Rev-erbα, ACSL4 and NCOA4 between HFHG+ H/R2 group and si-NC+ HFHG+ H/R group ( P>0.05). Compared with HFHG+ H/R2 group, the cell viability was significantly increased, the levels of LPO were decreased, and the expression of Rev-erbα, ACSL4 and NCOA4 was down-regulated in si-Rev-erbα+ HFHG+ H/R group ( P<0.05). Conclusions:Rev-erbα participates in the process of HFHG and H/R injury to cardiomyocytes by negatively regulating ferroptosis.

Objective To investigate the influence of body mass index(BMI)on individualized analgesic effect,inflammatory factors and safety of sufentanil in elderly patients after proximal femoral nail anti-rotation(PFNA)surgery.Methods A total of 161 elderly patients who received PFNA surgery in Nanjing Drum Tower Hospital Group Suqian Hospital from January 2022 to December 2023 were selected as study subjects.Patients with BMI<18.5 kg/m2 were set as Group A,patients with BMI ranging from 18.5 kg/m2 to 23.9 kg/m2 were set as Group B,patients with BMI ranging from 24.0 kg/m2 to 35.0 kg/m2 were set as Group C.After operation,individualized analgesia was performed with an intravenous patient-controlled analgesia pump of 2 μg/kg sufentanil based on body weight,and the postoperative pain degree,inflammatory factors,analgesia condition and adverse reactions of patients in the three groups were compared.Results At 8 hours,12 hours,24 hours and 48 hours after surgery,the pain visual analogue scale(VAS)scores of patietns in group B and group C were significantly lower than those in group A(P<0.05).At 12 hours,24 hours and 48 hours after surgery,the pain VAS scores of patients in group C were significantly lower than those in group B(P<0.05).The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)1 day after surgery of patients in group B and group C were significantly lower than those in group A(P<0.05).The pressing times of analgesia pump and the duration of obvious pain within 48 hours after surgery of patients in group A were significantly more/longer than those in group B and group C(P<0.05),and the analgesic satisfaction score was significantly lower than those in group B and group C(P<0.05).The total incidence of analgesic adverse reactions in group C was significantly higher than those in group A and group B(P<0.05).Conclusion BMI may affect the individualized analgesic effect,inflammatory factors and safety of sufentanil in elderly patients undergoing PFNA surgery.Underweight patients may have insufficient analgesia,while overweight or obese patients may have excessive analgesia,which may affect the analgesic safety.