Objective To investigate the effect of inhibition of glycogen synthase kinase?3 beta ( GSK?3β) activity on sevoflurane postconditioning?induced cardioprotection in diabetic rats. Methods Healthy adult male Sprague?Dawley rats, weighing 250-300 g, in which diabetes mellitus was induced by intraperitoneal 1% streptozotocin 60 mg∕kg combined with high?fat and high?sucrose diet and confirmed by blood glucose level >16. 7 mmol∕L. Forty rats with diabetes mellitus were divided into 5 groups ( n=8 each) using a random number table: sham operation group ( S group ) , ischemia?reperfusion ( I∕R ) group, sevoflurane postconditioning group ( SP group) , GSK?3β inhibitor SB216763 group ( SB group) , and sevoflurane postconditioning plus SB216763 group ( SS group ) . Myocardial ischemia was induced by 30 min occlusion of the left anterior descending branch of the coronary artery followed by 120 min reperfu?
sion. The rats inhaled sevoflurane with the end?tidal concentration of 2.5% for 5 min starting from 1 min be?fore reperfusion in group SP. SB216763 0.2 mg∕kg was injected via the caudal vein at 1 min before reperfu?sion in group SB. In group SS, the rats inhaled sevoflurane with the end?tidal concentration of 2.5% for 5 min starting from 1 min before reperfusion, and SB216763 0.2 mg∕kg was injected via the caudal vein at 1 min before reperfusion. At 120 min of reperfusion, blood samples were collected from the carotid artery for determination of serum creatine kinase?MB (CK?MB) activity and cardiac troponin I (cTnI) concentra?tions. Myocardial specimens were collected at 120 min of reperfusion for microscopic examination of the pathological changes and for determination of myocardial infarct size ( by 2,3,5?triphenyltetrazolium chlo?ride staining) and phosphorylated GSK?3β (p?GSK?3β) expression (by Western blot). Results Com?pared with group S, the myocardial infarct size and serum CK?MB activity and cTnI concentration were sig?nificantly increased, and the expression of p?GSK?3βwas significantly down?regulated in I∕R, SP, SB and SS groups (P<0.05). Compared with group I∕R, the myocardial infarct size and serum CK?MB activity and cTnI concentration were significantly decreased, and the expression of p?GSK?3β was significantly up?regulated in SB and SS groups (P<0.05), and no significant change was found in the parameters men?tioned above in group SP ( P>0.05) . Compared with group SB, the myocardial infarct size and serum CK?MB activity and cTnI concentration were significantly decreased, and the expression of p?GSK?3β was sig?nificantly up?regulated in group SS (P<0.05). The pathological changes of myocardium were significantly attenuated in SB and SS groups as compared with group I∕R and group SP . Conclusion Inhibition of GSK?3β activity can improve sevoflurane postconditioning?induced cardioprotection in diabetic rats.

TNFR-Fc is an important fusion protein that has great potential in therapeutic and diagnostic applications. We developed an efficient fed-batch process for GS-CHO cells to produce TNFR-Fc. The rationale of this fed-batch process relies on the supply of sufficient nutrients to meet the requirements of cell metabolism. The optimal feed medium was designed through ration design. A metabolically responsive feeding strategy was designed and dynamically adjusted based on the residual glucose concentration determined off-line. In this process, the maximal viable cell density and antibody concentration reached above 9.4x10(6) cells/mL and 207 mg/L, respectively. Compared with the batch process, the newly developed fed-batch process increased the cell yield by 3.4 fold and the final antibody concentration by 3 fold. This fed-batch process would therefore facilitate the production of therapeutic antibody by GS-CHO cells.
Animals ; CHO Cells ; Cell Culture Techniques ; methods ; Cricetinae ; Cricetulus ; Culture Media ; Etanercept ; Glucose ; analysis ; Immunoglobulin G ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics

Objective:To investigate the comparative analysis of multimodal ultrasound features and clinicopathology of different molecular types of breast cancer, so as to improve the diagnostic accuracy of different types of breast cancer.Methods:A retrospective analysis of 112 patients with invasive breast cancer in the Second Affiliated Hospital and Affiliated Cancer Hospital of Harbin Medical Univeristy from August 2017 to November 2018 was carried out. With reference to St.Gallen standard, breast masses were divided into four subtypes, including Luminal A (LA), Luminal B (LB), Her-2 overexpression (Her-2+ ) and triple negative (TN). Two-dimensional greyscale, color Doppler flow imaging, shear wave elastography and contrast-enhanced ultrasound images were performed before operation. Complete clinicopathological data were collected. The differences between different subtypes of breast cancer in terms of multimodal ultrasound and clinicopathological features were analyzed.Results:①Luminal breast cancer showed posterior echo attenuation, low Alder grade, low Emax and Eratio value. LA was more likely to show a speculated or angular margin and concentric enhancement; LB was more frequently to show a indistinct margin, low enhancement and no perfusion defect (all P<0.05). ②Her-2+ cancers were characterized with a indistinct margin, enhanced posterior echo, a high Alder grade, Emax and Eratio value, and a high contrast enhancement and perfusion defect. The TTP value was lower than other subtypes (all P<0.05). ③TN cancers were associated with microlobulated margin, posterior echo enhancement, low Alder grade, high Emax and Eratio value and Ki67 expression, contrast concentric enhancement have and perfusion defect (all P<0.05). Conclusions:Different molecular subtypes of breast cancer have different multimodal ultrasound features and clinicopathological appearances, which can provide more imaging information for clinical diagnosis, treatment, and prognosis.

Objective:To construct a quantitatively diagnostic nomogram model by analyzing the clinical information of patients and the features of multi-modality ultrasound images of thyroid lesions, so as to preoperatively predict the malignant probability of suspicious thyroid nodules and provide effective references for clinical decision-making.Methods:A total of 933 patients, 1 121 thyroid nodules of C-TIRADS 3-5 categories, who underwent surgery in the Second Affiliated Hospital of Harbin Medical University from September 1, 2020 to June 10, 2021 were collected. The nodules were randomly divided into training ( n=897) and test groups ( n=224) in 8∶2 ratio. Finally, the diagnostic performance was evaluated by area under the curve (AUC). Results:①After preliminary screening by univariate analysis, multivariate analysis showed that age, echogenicity, orientation, echogenic foci, margin, posterior features, and elastic score were significantly correlated with benign and malignant nodules (all P<0.001), and the difference of halo between benign and malignant nodules was also statistically significant ( P=0.012). ②The AUC of nomogram was up to 0.903(95% CI=0.862-0.944) in the test set, and 0.889(95% CI=0.832-0.946) and 0.960(95% CI=0.925-0.994) in nodules with maximum diameter of ≤10 mm and of >10 mm respectively, which showed high diagnostic performance. Conclusions:The nomogram model could accurately differentiate malignant from benign thyroid nodules preoperatively, with the highest diagnostic performance for the nodules with maximum diameter of >10 mm, and effectively avoid the unnecessary fine-needle biopsy and surgical operation.

Objective:To explore the correlation between tumor markers and prognosis of patients with idiopathic inflammatory myopathy (IIM) associated interstitial lung disease (ILD).Methods:A total of 149 patients who were no less than 18 years old and diagnosed with IIM-ILD from July 2017 to September 2019 in the First Affiliated Hospital of Zhengzhou University were consecutively enrolled in the study. Ten patients were lost to follow-up. The remaining 139 cases were regarded as research objects. Patients were divided into survival group or death group according to their one-year survival status. Then their baseline characteristics were compared. Univariate Cox regression analyses of age, gender, cancer, inflammatory indexes, muscle zymogram, tumor markers, ferritin, melanoma differentiation-associated gene 5 (MDA5) antibody and treatment regimens were conducted to identify prognostic risk factors of one-year mortality. Corrected multivariable cox regression was applied to screen the independent risk factors associated with one-year mortality of IIM-ILD. According to the cut-off value of carcinoembryonic antigen (CEA) and neuron specific enolase (NSE) (6 μg/L and 28 μg/L, respectively), patients were divided into high-level groups and low-level groups. Kaplan Meier survival curve were generated to compare one-year survival rate of high-level groups and low-level groups. On the basis of qualitative results of MDA5 antibody, patients were split into two groups with positive MDA5 antibody or negative MDA5 antibody. The differences of CEA, NSE levels between the two groups and the correlation between CEA, NSE levels and ferritin were analyzed.Results:Age, lactate dehydrogenase (LDH), CEA, carbohydrate antigen (CA) 199, NSE and ferritin in the death group were higher than those in the survival group, while the rate of immunosuppressant administration was lower than that in survival group ( P<0.05). Univariate regression analyses showed that CEA, cytokeratin 19 fragment (CYFRA211) and NSE were risk factors for one-year mortality of IIM-ILD. Adjusted by age, treatment regimens and tumor, multivariate regression analysis showed that CEA [ HR=1.112, 95% CI (1.017-1.214), P=0.019] and NSE [ HR=1.033, 95% CI (1.002-1.064), P=0.034] were independent risk factors for one-year mortality. One-year survival rate of the group with CEA≥6 μg/L was lower than that in the group with CEA<6 μg/L (Logrank test, P<0.001). Similarly, one-year survival rate of the group with NSE≥28 μg/L was lower than that in the group with NSE<28 μg/L (Logrank test, P<0.001). In addition, the CEA level in patients with positive MDA5 antibody was higher than that in patients with negative MDA5 antibody ( P<0.001). However, there was no correlation between NSE and MDA5 antibody. Moreover, serum levels of CEA ( r=0.299, P=0.002) and NSE ( r=0.349, P<0.001) were positively correlated with ferritin. Conclusions:Tumor markers have predictive value for the prognosis of IIM-ILD. Higher CEA and NSE are independent risk factors for poor prognosis in patients with IIM-ILD.

Objective To analyze the radiation exposure of medical personnel, and to provide real-time radiation dose monitoring data for medical personnel in interventional surgeries. Methods A total of 96 person-times who participated in cardiac surgeries in interventional operating rooms from September 2019 to March 2022 were selected as subjects. The subjects were divided into two groups according to the time progress, with 43 person-times in intervention group and 53 person-times in control group. The subjects in the intervention group were provided with radiation dose monitoring data during surgeries, while the subjects in the control group performed surgeries with routine procedures. The average intraoperative radiation dose rates of the two groups were compared. Results The median radiation dose rate was significantly lower in the intervention group than in the control group (88.80 μSv/h vs 188.40 μSv/h, U = 637.000, P = 0.000). Conclusion Providing real-time radiation dose monitoring data for medical personnel in interventional surgeries can effectively reduce their occupational radiation exposure and improve their occupational radiation protection.

ObjectiveTo explore the safety and efficacy of robot-assisted minimally invasive esophagectomy （robot-assisted minimally invasive esophagectomy， RAMIE） and thoracic laparoscopy combined with minimally invasive esophageal resection （minimal invasive esophagectomy， MIE）. MethodsThe data of 188 patients treated with Da Vinci robot assisted minimally invasive esophageal resection （RAMIE） from April 2021 to December 2022 were analyzed. In the RAMIE group， 69 patients， 49 males and 20 female， age （67.2 ± 7.2）； 119 in the MIME group， respectively， 89 males and 30 female， age （69.1 ± 7.0）. At 1 ∶ 1， including 58 patients in the RAMIE group and 58 patients in the MIE group. The t-test， Wilcoxon rank-sum test， χ² test， and so on. ResultsAfter PSM treatment， the clinical data between the two groups. There was no significant difference in operation time， postoperative tube days， and total number of lymph node dissection between the RAMIE and MIE groups （P <0.05）； the RAMIE group was better in terms of intraoperative bleeding and the MIE group， statistically significant （P <0.05）； the MIE group was better in drainage flow and lymph node dissection for three days （P <0.05）. In terms of postoperative complications， there was no statistical difference between RAMIE and MIE groups （P>0.05）. ConclusionThe recent efficacy of robot-assisted minimally invasive esophagectomy is comparable to that of thoracic laparoscopy and minimally invasive Mckeown esophagectomy； robotic-assisted minimally invasive esophagectomy can reduce intraoperative bleeding and have more advantages in left recurrent laryngeal nerve lymph node dissection.

The aim of this study is to quantify the 146S antigen in foot-and-mouth disease virus (FMDV) inactivated vaccine by size-exclusion chromatography (SEC). The analysis was performed on a TSKgel G4000SWXL column (7.8 mm×30 cm), with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV (serotype O) 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL (R2=0.996, n=10), and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6% (RSD=2.7%, n=3), 102.3% (RSD=2.6%, n=3), and 95.5% (RSD=5.1%, n=3). The method was proved accurate and reliable with good reproducibility (RSD=0.5%, n=6), and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control.