To investigate the role of remote limb ischemic postconditioning in mitigation of focal cerebral ischemia-reperfusion injury in rats via mitochondrial autophagy.

Totally 105 adult male Sprague-Dawley rats were divided into a sham operation group, a ischemia-reperfusion group (A group), a ischemia-reperfusion + remote ischemic postconditioning group (B group), a ischemia-reperfusion + remote ischemic postconditioning + isotonic NaCl solution group (C group) and a ischemia-reperfusion + remote ischemic postconditioning + mitochondrial division inhibitor-1 (Mdivi-1) group (D group), 21 rats in each group. The right carotid artery was exposed and freed in the sham operation group, while a model of focal cerebral ischemia-reperfusion injury was prepared by middle cerebral artery blockade in the A, B, C and D groups. Then 10 min of ischemia/10 min of reperfusion were performed for three cycles in the B, C and D groups, and an equal volume of isotonic NaCl solution and 3 mg/kg of Mdivi-1 were injected intraperitoneally in the C and D groups 5 min before ischemia respectively. The neurological deficit score (NDS), percentage of cerebral infarction volume, and apoptosis rate, microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ/Ⅰ ratio, superoxide dismutase (SOD), malondialdehyde and 15-F_2t-Isoprostane of cerebral ischemic semidark cells were compared.

No neurological deficit and cerebral infarction occurred in the sham operation group. The NDS [(2.8 ± 0.6), (1.6 ± 0.4), (1.6 ± 0.5), (2.5 ± 0.5) scores] and percentage of cerebral infarction volume [(48 ± 3)%, (28 ± 4)%, (28 ± 4)%, (41 ± 3)%] were statistically significantly different in the A, B, C and D groups (F = 39.237, 53.278; both P < 0.001). Moreover, they were significantly lower in the B and C groups than in the A and D groups (all P < 0.05). The apoptosis rate [(2.3 ± 0.8)%, (54.6 ± 5.2)%, (29.3 ± 3.1)%, (29.8 ± 3.3)%, (51.2 ± 4.5)%], LC3-Ⅱ/Ⅰ ratio [(0.13 ± 0.03), (0.32 ± 0.05), (0.53 ± 0.06), (0.48 ± 0.08), (0.35 ± 0.06)], SOD [(168 ± 19), (92 ± 13), (162 ± 21), (165 ± 23), (94 ± 15) U/mg], malondialdehyde [(4.22 ± 0.28), (8.41 ± 0.42), (5.14 ± 0.27), (5.26 ± 0.31), (7.93 ± 0.44) nmol/mg] and 15-F_2t-Isoprostane [(179 ± 86), (389 ± 105), (208 ± 89), (215 ± 85), (364 ± 103) mg/g] of cerebral ischemic semidark cells were statistically significantly different in the sham operation, A, B, C and D groups (F = 54.658, 32.358, 59.677, 46.195, 193.962; all P < 0.001). Further pairwise comparison showed that the apoptosis rate, LC3-Ⅱ/Ⅰ ratio, malondialdehyde and 15-F_2t-Isoprostane in the A, B, C and D groups were significantly higher than those in the sham operation group, while the SOD expression level in the A and D groups was significantly lower than that in the sham operation group (all P < 0.05). Compared with the A and D groups, the LC3-Ⅱ/Ⅰ ratio and SOD expression level of rats in the B and C groups significantly increased, and the apoptosis rate, malondialdehyde and 15-F_2t-Isoprostane significantly decreased (all P < 0.05).

Remote limb ischemia postconditioning may reduce focal cerebral ischemia-reperfusion injury in rats by increasing the mitochondrial autophagy level and inhibiting the oxidative stress response.

To investigate the effect of resveratrol on the nuclear factor-E2 related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) signaling pathway in myocardial ischemia-reperfusion (I/R) injury of type 2 diabetes mellitus rats.

Sixty Sprague-Dawley rats with type 2 diabetes mellitus were randomly divided into a sham operation group, a I/R group, a resveratrol preconditioning (R) group, and a resveratrol preconditioning + EX527 (RE) group, 15 rats in each group. Myocardial I/R injury was produced by ligating the left coronary anterior descending artery for 30 min followed by 120 min reperfusion. Rats in the R group and RE group were intraperitoneally injected with resveratrol (15 mg/kg, once a day) for 7 days before the operation; rats in the sham operation group and I/R group were intraperitoneally injected with the equal capacity of isotonic NaCl solution; rats in the RE group also received EX527 (1 μg/kg) by caudal vein injection at 15 min before ischemia. At the end of 120 min reperfusion, blood samples were taken for detecting the concentration of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) by the enzyme-linked immunosorbent assay. Then rats were sacrificed in each group; the pathological changes of myocardium were observed by hematoxylin-eosin (HE) staining and the myocardial infarct size was measured by 2,3,5-triphenyltetrazolium chloride. The levels of malondialdehyde and superoxide dismutase (SOD) were compared among the four groups. The expression levels of myocardial silent information regulator of transcription 1 (SIRT1), Nrf2, and HO-1 protein were detected using Western-blotting.

From HE staining, the myocardial cells were slightly broken and edematous, with a small amount of inflammatory cells in the sham operation group; the myocardial fibers were disorderly arranged, with myocardial interstitial congestion and edema infiltrated by a large number of inflammatory cells in the I/R group; the myocardial tissue was relatively structurally neat, with occasional karyopyknosis in the R group; the myocardial fibers were arranged in a disorderly way, with obvious karyopyknosis and inflammatory cells in the RE group. No myocardial infarction occurred in the sham operation group. The myocardial infarct size in the I/R group, R group, and RE group showed significant differences [(51 ± 6)%, (37 ± 4)%, (41 ± 3)%; F = 160.703, P < 0.001]. It was much smaller in the R group and RE group than in the I/R group, and was smallest in the R group (all P < 0.05). The levels of LDH, CK-MB, malondialdehyde, SOD, SIRT1, Nrf2, and HO-1 all showed significant differences among the four groups (F = 144.101, 158.545, 53.682, 99.273, 50.121, 59.153, 143.702; all P < 0.001). Compared with the sham operation group, the levels of LDH, CK-MB, and malondialdehyde increased obviously in the I/R group, R group and RE group, and the levels of SOD, SIRT1, Nrf2, and HO-1 decreased (all P < 0.05). Compared with the I/R group, the levels of LDH, CK-MB, and malondialdehyde decreased obviously in the R group and RE group, which decreased most in the R group, and the levels of SOD, SIRT1, Nrf2, and HO-1 increased, which increased most in the R group (all P < 0.05).

Resveratrol can alleviate oxidative stress and myocardial I/R injury in type 2 diabetes mellitus rats by promoting SIRT1 to activate the Nrf2/HO-1 signaling pathway.

To study risk factors for the low cardiac output syndrome (LCOS) after total anomalous pulmonary venous connection (TAPVC) correction.

Totally 153 child patients undergoing TAPVC correction were selected from the Center of Pediatric Cardiology of Beijing Anzhen Hospital, Capital Medical University between January 2014 and January 2018. They were divided into a LCOS group (n = 50) and a no LCOS group (n = 103) according to whether the LCOS appeared after operation. Clinical data before, during and after operation were compared between the two groups, and risk factors of LCOS after TAPVC correction were analyzed by Logistic regression analysis.

The perioperative mortality of child patients in the LCOS group was significantly higher than that in the no LCOS group [22.0% (11/50) vs. 3.9% (4/103), χ² = 12.493, P < 0.001]. The age [3 (1, 5) months vs. 5 (2, 12) months, H = 2.722, P = 0.006], body mass [5.2 (4.5, 6.0) kg vs. 6.0 (5.0, 8.0) kg, H = 3.519, P < 0.001], preoperative left ventricular end-diastolic diameter [15 (13, 17) mm vs. 18 (15, 23) mm, H = 4.170, P < 0.001], preoperative atrial septal defect size [6 (4, 8) mm vs. 8 (6, 11) mm, H = 3.368, P = 0.001], preoperative oxygen saturation [85 (80, 86)% vs. 85 (82, 87)%, H = 2.168, P = 0.030], intraoperative cardiopulmonary bypass time [100 (75, 137) min vs. 88 (70, 109) min, H = 2.459, P = 0.014] and delayed chest closure (χ² = 4.484, P = 0.034) were statistically significantly different between the LCOS group and no LCOS group. Then the age, body mass, preoperative left ventricular end-diastolic diameter, preoperative atrial septal defect size, preoperative oxygen saturation, delayed chest closure and intraoperative cardiopulmonary bypass time were included in the Logistic regression analysis. The results showed that the preoperative left ventricular end-diastolic diameter [odds ratio (OR) = 0.851, 95% confidence interval (CI) (0.732, 0.989), P = 0.035] and preoperative oxygen saturation [OR = 0.901, 95%CI (0.829, 0.979), P = 0.014] were protective factors of LCOS after TAPVC correction, and the intraoperative cardiopulmonary bypass time [OR = 1.012, 95%CI (1.001, 1.022), P = 0.028] was its risk factor.

Attention should be paid to the preoperative left ventricular end-diastolic diameter, oxygen saturation and intraoperative cardiopulmonary bypass time of TAPVC patients. If LCOS signs are found, early intervention should be made to improve their prognosis.

To establish a nomogram for early prediction of the severity of patients with coronavirus disease 2019 (COVID-19) to guide clinical treatment.

From January 17 to February 14, 2020, 116 patients with COVID-19 admitted to Wenzhou Central Hospital were selected. According to their clinical manifestations, 116 patients were divided into a mild group (n = 4), a common group (n = 90), a severe group (n = 18) and a critical group (n = 4). The hospitalization time and complications of all patients were recorded, and the general data and clinical indicators were compared among the 4 groups. Risk factors for the prognosis of patients with COVID-19 were obtained by multivariate Logistic regression analysis. Then a visual regression nomogram was established using R language software. Finally, efficacy of the nomogram was detected by a receiver operating characteristic (ROC) curve.

The age [(39 ± 11), (43 ± 12), (53 ± 13), (60 ± 8) years; F = 5.815, P = 0.001], C-reactive protein [1.7 (0.6, 7.1), 7.9 (2.9, 21.6), 28.4 (13.9, 42.5), 61.7 (44.7, 79.8) mg/L; H = 8.424, P < 0.001], hematocrit [(36 ± 5)%, (41 ± 4)%, (39 ± 4)%, (37 ± 5)%; F = 4.344, P = 0.006], platelet count [318.0 (251.0, 409.0) × 10⁹/L, 180.5 (140.0, 225.5) × 10⁹/L, 162.0 (130.0, 222.8) × 10⁹/L, 108.5 (82.0, 103.0) × 10⁹/L; H = 7.225, P < 0.001], aspartate aminotransferase [16.0 (15.5, 19.5), 23.0 (19.0, 31.0), 34.5 (26.3, 55.0), 39.5 (29.0, 82.3) U/L; H = 6.159, P = 0.001], albumin [(44 ± 6), (43 ± 16), (39 ± 3), (33 ± 4) g/L; F = 9.508, P < 0.001], and lactate dehydrogenase [142.5 (107.8, 189.3), 198.0 (159.5, 238.0), 295.0 (251.0, 323.0), 369.5 (295.2, 436.3) U/L; H = 14.225, P < 0.001] of patients with COVID-19 in the 4 groups were statistically significantly different. Then the age, C-reactive protein, hematocrit, platelet count, aspartate aminotransferase, albumin, and lactate dehydrogenase were incorporated into multivariate Logistic regression analysis. It showed that the albumin [odds ratio (OR) = 0.756, 95% confidence interval (CI) (0.581, 0.982), P = 0.036] and lactate dehydrogenase [OR = 1.019, 95%CI (1.007, 1.032), P = 0.002] were risk factors for the prognosis of patients with COVID-19. In the meantime, a reliable and intuitive nomogram was obtained by R language software. ROC curve analysis showed that this nomogram had excellent predictive ability for severe and critical patients [area under the curve (AUC) = 0.903, 95%CI (0.831, 0.975), P < 0.001], for critical patients alone [AUC = 0.974, 95%CI (0.932, 1.000), P < 0.001] and for severe patients alone [AUC = 0.848, 95%CI (0.759, 0.937), P < 0.001].

This nomogram can predict the severity of patients with COVID-19 early and effectively, and may be a practical tool to guide clinical treatment.

To explore the application value of CT-guided percutaneous needle biopsy assisted by the IG4 electromagnetic navigation system in diagnosing pulmonary lesions.

A total of 125 patients with pulmonary lesions to undergo CT-guided percutaneous needle biopsy were randomly divided into a control group (62 patients) and a magnetic group (63 patients). Patients in the control group were performed conventional CT-guided percutaneous needle biopsy, and patients in the magnetic group received CT-guided percutaneous needle biopsy assisted by the IG4 electromagnetic navigation system. The white blood cell count, platelet count (PLT), international normalized ratio (INR), activated partial thromboplastin time (APTT), prothrombin time (PT), alanine aminotransferase (ALT), serum creatinine, creatine kinase-MB (CK-MB), C-reactive protein (CRP), and lung cancer related markers of neuronspecific enolase (NSE) and serum squamous cell carcinoma antigen (SCCA) before the piercing were recorded. The positioning time, needle adjustment times, scanning times, radiation dose [dose length product (DLP)], pathological diagnosis sensitivity and incidence of complications such as pneumothorax and bleeding were compared between the two groups.

The levels of white blood cell count, PLT, INR, APTT, PT, ALT, serum creatinine, CK-MB, CRP, NSE and SCCA before puncture all showed no significant differences between the two groups (all P > 0.05). Meanwhile, the positioning time [(9.5 ± 2.6) min vs. (13.6 ± 4.1) min, t = 6.584, P < 0.001], needle adjustment times [(3.0 ± 1.3) times vs. (3.8 ± 1.8) times, t = 2.662, P = 0.009], scanning times [(3.9 ± 1.2) times vs. (5.8 ± 1.8) times, t = 6.853, P < 0.001] and DLP [(291 ± 167) mGy × cm vs. (407 ± 307) mGy × cm, t = 2.622, P = 0.010] in the magnetic group were significantly better than those in the control group. The incidence of pneumothorax [15.87% (10/63) vs. 33.87% (21/62), χ² = 5.204, P = 0.023] and bleeding [20.63% (13/63) vs. 38.71% (24/62), χ² = 4.899, P = 0.027] in the magnetic group was significantly lower than that in the control group. However, there was no statistically significant difference in the pathological diagnosis sensitivity between the two groups [89.66% (52/58) vs. 96.36% (53/55), χ² = 1.931, P = 0.165].

The CT-guided percutaneous needle biopsy assisted by the IG4 electromagnetic navigation system is significantly superior to conventional CT-guided lung biopsy in terms of safety and degree of injury to patients, and has high clinical application value.

To establish a multiplex fluorescent PCR-melting curve method for simultaneous detection of Candida, Aspergillus and Cryptococcus, and to evaluate its diagnostic value.

A multiplex fluorescent PCR-melting curve method was established and its detection system was optimized. Totally 179 clinical specimens including 65 blood specimens, 35 deep sputum specimens, 30 urine specimens, 18 cerebrospinal fluid specimens, 12 hydrothorax and ascites specimens, 10 alveolar lavage fluid specimens and 9 fresh lung tissue specimens for highly suspected invasive fungal infection (IFI) were enrolled in this study. The sensitivity, specificity and repeatability experiments were used to verify the detection performance of this method, and the receiver operating characteristic (ROC) curve was used to evaluate its diagnostic efficiency.

The multiplex fluorescent PCR-melting curve method could simultaneously detect 8 common pathogenic fungi, including 4 kinds of Candida (Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei), 3 kinds of Aspergillus (Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger) and Cryptococcus neoformans. The minimum detection limit was 1 × 10⁴ cfu/mL. Common bacteria had no amplification reaction, and the melting temperature fluctuation of repetitive experiment was less than 0.5 ℃. Among 179 clinical specimens, 112 were positive by "gold standard" methods such as culture, microscopy and pathological diagnosis, and 96 were positive by the multiplex fluorescent PCR-melting curve method. The sensitivity, specificity, positive predictive value and negative predictive value for simultaneous detection of Candida, Aspergillus and Cryptococcus neoformans by this method were 0.857, 0.970, 0.980 and 0.802, respectively. The area under ROC curve was 0.914, and the 95% confidence interval was 0.868 ~ 0.959 (P < 0.001).

The multiplex fluorescent PCR-melting curve method can detect Candida, Aspergillus and Cryptococcus neoformans rapidly and accurately with high throughput, which is of great significance for the early diagnosis of IFI.