Public cell culture platform is an important facility in laboratory animal facilities, providing essential support for scientific research such as the development of animal tumor disease models and transgenic animals. By establishing a public cell culture experimental platform, laboratory animal centers can effectively integrate experimental animals and cell culture resources, optimizing the allocation of scientific research resources to facilitate better research outcomes. The majority of cells cultured in these platforms are used for animal experiments. Contamination or quality issues in these cells not only affect experimental results but also jeopardize the health of experimental animals, potentially leading to microbial infections and contamination of entire animal facilities. Therefore, public cell culture laboratories within experimental animal facilities impose stricter quality control measures than conventional cell culture rooms. This study takes the public cell culture platform at the Laboratory Animal Center of Shanghai Jiao Tong University as a case study to discuss management experiences, focusing on facility maintenance and management, personnel management and quality control of cell biological risk. The aim is to provide useful reference for the management of public cell culture laboratories in experimental animal facilities and other institutions.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Objective:To explore the mediating effect of appearance functional internalization between social comparison tendency and body image distress of college students.Methods:From June to September 2023, a cross-sectional survey was conducted among 308 college students with the social comparison tendency scale, the functional internalization questionnaire of college students' appearance and the body image anxiety scale for adolescent students.SPSS 22.0 software was used for common method bias test, descriptive statistics and correlation analysis, Mplus 8.0 was used for structural equation construction, and Bootstrap method was used for intermediary effect analysis.Results:Body image distress (43.86±8.78) was positively correlated with social comparison tendency (35.20±6.04) and appearance functional internalization (38.35±9.68) ( r=0.35, 0.33, both P<0.01).There was also a positive correlation between social comparison tendency and appearance functional internalization ( r=0.51, P<0.01). The effect of social comparison tendency on body image distress was mediated by appearance functional internalization, and the mediating effect value was 0.137, accounted for 35.13%(0.137/0.390) of the total effect. Conclusion:The appearance functional internalization mediates the relationship between social comparison tendency and body image distress.Social comparison tendency can not only directly predict body image distress, but also can predict body image distress, indirectly through appearance functional internalization.

Objective:To explore the effect of joint segmentation model of myocardial-fibrotic region based on deep learning in quantitative analysis of myocardial fibrosis in patients with dilated cardiomyopathy(DCM).Methods:The data of 200 patients with confirmed DCM and myocardial fibrosis in the left ventricle detected by cardiac MR-late gadolinium enhancement (CMR-LGE) in Xuzhou Central Hospital from January 2015 to April 2022 were retrospectively analyzed. Using a complete randomized design, the patients were divided into training set ( n=120), validation set ( n=30) and test set ( n=50). The left ventricle myocardium was outlined and the normal myocardial region was selected by radiologists. Fibrotic myocardium was extracted through calculating the threshold with standard deviation (SD) as a reference standard for left ventricle segmentation and fibrosis quantification. The left ventricular myocardium was segmented by convex prior U-Net network. Then the normal myocardial image block was recognized by VGG image classification network, and the fibrosis myocardium was extracted by SD threshold. The myocardial segmentation effect was evaluated using precision, recall, intersection over union (IOU) and Dice coefficient. The consistency of myocardial fibrosis ratio in left ventricle obtained by joint segmentation model and manual extraction was evaluated with intra-class correlation coefficient (ICC). According to the median of fibrosis rate, the samples were divided into mild and severe fibrosis, and the quantitative effect of fibrosis was compared by Mann-Whitney U test. Results:In the test set, the precision of myocardial segmentation was 0.827 (0.799, 0.854), the recall was 0.849 (0.822, 0.876), the IOU was 0.788 (0.760, 0.816), and the Dice coefficient was 0.832 (0.807, 0.857). The consistency of fibrosis ratio between joint segmentation model and manual extraction was high (ICC=0.991, P<0.001). No statistically significant difference was found in the ratio error between mild and severe fibrosis ( P>0.05). Conclusions:The joint segmentation model realizes the automatic calculation of myocardial fibrosis ratio in left ventricle, which is highly consistent with the results of manual extraction. Therefore, it can accurately realize the automatic quantitative analysis of myocardial fibrosis in patients with dilated cardiomyopathy.

OBJECTIVES: Sepsis associated encephalopathy (SAE) is a common neurological complication of sepsis. Delirium is a common symtom of SAE. The pathophysiology of SAE is still unclear, but several likely mechanisms have been proposed, such as mitochondrial and endothelial dysfunction, neurotransmission disturbances, derangements of calcium homeostasis, cerebral microcirculation dysfunction, and brain hypoperfusion. Near-infrared spectroscopy (NIRS) is a non-invasive measure for regional cerebral oxygen saturation (rSO METHODS: A total of 48 septic patients who admitted to the intensive care unit (ICU) of Xiangya Hospital, Central South University from August 2017 to May 2018, were retrospectively study. Septic shock was diagnosed according to the criteria of sepsis 3.0 defined by the American Association of Critical Care Medicine and the European Society of Critical Care Medicine. NIRS monitoring was performed during the first 6 hours admitted to ICU with sensors placed on the bilateral forehead of patients. The maximum (rSO RESULTS: The overall 28-day mortality of septic shock patients was 47.92% (23/48), and the incidence of delirium was 18.75% (9/48). The rSO CONCLUSIONS Cerebral anoxia and hyperoxia, as well as the large fluctuation of cerebral oxygen saturation are important factors that affect the outcomes and the incidence of delirium in septic shock patients, which should be paid attention to in clinical practice. Dynamic monitoring of cerebral oxygen saturation and maintain its stability may be of great significance in patients with septic shock.
APACHE ; Humans ; Intensive Care Units ; Oxygen Saturation ; Prognosis ; Retrospective Studies ; Sepsis ; Shock, Septic

Objective:To explore the correlation between different types of microcirculation alterations and the prognosis in patients with septic shock.Methods:This research employed a prospective observational study methodology for selecting subjects with septic shock. Side-stream dark field(SDF) was used to monitor the sublingual microcirculation to determine the total vascular density (TVD), perfused vessel density (PVD), the proportion of perfused vessels (PPV), and the microvascular flow index (MFI), heterogeneity index (HI) indicators. At the bedside, patients with microcirculation disorders were divided into four types: stasis, dilution, heterogeneity, and hyperdynamic. The 30-day survival status after enrollment and hemodynamics parameters were recorded.Results:A total of 64 patients with septic shock were selected in the study, including 18 cases of stasis type, 11 of dilution type, 18 of heterogeneous type, and 17 of hyperdynamic type. There were statistical differences in the mean arterial pressure (MAP) [stasis:(77±9) mmHg (1 mmHg=0.133 kPa), dilution:(80±11) mmHg, heterogeneity: (78±12) mmHg, hyperdynamic:(88±12) mmHg], TVD [ stasis:(10.84±3.01) mm/mm 2, dilution:(9.64±1.72) mm/mm 2, heterogeneity:(11.39±2.18) mm/mm 2, hyperdynamic: (11.87±2.67) mm/mm 2 ], PVD [stasis:(5.93±1.94) mm/mm 2, dilution:(6.86±1.48) mm/mm 2, heterogeneity: (8.31±1.78) mm/mm 2, hyperdynamic:(9.68±2.46) mm/mm 2], PPV [stasis:52.45 (46.25, 63.33)%, dilution:73.70 (61.50, 75.20)%, heterogeneity: 71.25 (67.95, 77.00)%, hyperdynamic:80.70 (77.25, 86.45)%], MFI(stasis:1.34±0.45, dilution: 1.70±0.38, heterogeneity:1.82±0.28, hyperdynamic:2.25±0.33), and HI [stasis:0.68 (0.51, 1.87), dilution: 0.57 (0.49, 0.64), heterogeneity:0.70 (0.59, 0.91), hyperdynamic: 0.40 (0.37, 0.52)] of the four types of microcirculation alterations. The cumulative survival rates in stasis, dilution, heterogeneity and hyperdynamic types at 30 day were 7/18, 4/11, 10/18 and 14/17, respectively, which in stasis and dilution types was significantly lower than that of hyperdynamic type (χ2=7.221, P=0.007;χ 2=6.764, P=0.009). Multivariate Cox regression analysis showed the type of microcirculation alterations (stasis: RR=4.551, 95 %CI 1.228-16.864, P=0.023; dilution: RR=4.086, 95 %CI 1.011-16.503, P=0.048), acute physiology and chronic health evaluation Ⅱ ( RR=1.077, 95 %CI 1.006-1.153, P=0.032) were independent prognostic risk factors. Conclusions:Microcirculation alterations are common in patients with septic shock, and it is hard to predict the types of microcirculation alterations with hemodynamics parameters. The prognosis of patients with septic shock is related to the types of microcirculation alterations, suggesting that routine monitoring of microcirculation might be helpful to guide hemodynamic therapy.

Objectives:To study the feasibility of using ultrasound to evaluate diaphragm function in patients with invasive mechanical ventilation.Methods:From March to December 2017, 40 adult patients with acute respiratory distress syndrome who were admitted to the Department of Critical Care Medicine, Xiangya Hospital, Central South University for more than 48 hours were included. Diaphragmatic excursion and thickness of bilateral anterior, middle and posterior parts were measured by ultrasound for 5 consecutive days.Results:(1) Compared with the diaphragmatic excursion of the right [anterior: (11.05±3.04) mm; middle: (12.08±2.71) mm; posterior: (11.51±3.33) mm] and left [anterior: (13.63±7.52) mm; middle: (15.44±7.52) mm; posterior: (14.76±6.93) mm] sides on day 1, the diaphragmatic excursion of the right [anterior: (8.90±3.65) mm; middle: (10.02±4.24) mm; posterior: (10.25±4.38) mm] and left [anterior: (9.82±1.96) mm; middle: (11.60±1.13) mm; posterior: (11.52±1.98) mm] sides decreased significantly on day 3 ( P<0.05). Bilateral anterior, middle and posterior diaphragmatic excursion recovered on day 5, and was higher than the baseline levels on day 1, with the left middle and posterior diaphragmatic excursion changing most significantly. (2) Compared with day 1, 2, 3, the thickening fraction of bilateral anterior, middle and posterior diaphragm were significantly decreased on day 4, with the left middle part [day 1: (33.87±14.34)%; day 2: (37.26±13.91)%; day 3: (30.56±14.27)%; day 4: (15.53±5.68)%] and the left posterior part [day 1: (35.50±15.69)%; day 2: (39.84±15.32)%; day 3: (29.06±14.96)%; day 4: (13.30±5.79)%] changing most significantly ( P<0.05). The thickening fractions of left anterior, middle and posterior diaphragm recovered on day 5 compared with that on day 4, but still lower than those on day 1 ( P<0.05). Conclusions:It is feasible to evaluate the diaphragm function in patients with invasive mechanical ventilation by ultrasound, which can provide guidance for preventing diaphragmatic atrophy and withdrawing from mechanical ventilation.

Objective To evaluate the role of combining relative alpha variability and electroencephalogram (EEG) reactivity to predict the prognosis of hypoxic?ischemic encephalopathy(HIE) in adult patients. Methods A total of 28 adult patients with HIE admitted to general intensive care unit at Xiangya Hospital in Central South University were enrolled in this observational study from January2016 to April 2017. These patients with body temperature over 35℃after 72?hour admission could be continuously monitored at least 12 hours byEEG.At the same time,each patient was assessed for EEG reactivity.Then we analyzed the correlation between EEG reactivity, relative alpha variability and clinical prognosis. Results EEG reactivity was elicited in 15/28 patients, among whom 12 patients had a good outcome. While in the other 13 patients, EEG reactivity was not elicited, among whom only 3 patients had a good outcome. As to the results ofrelative alpha variability,11/13 patients with degree 3?4were of good prognosis; while only 3/15 patients with degree 1?2 were of good prognosis. Glasgow coma scale(GCS), EEG reactivity, and relative alpha variability were correlated with clinical outcome(χ2=5.073,9.073,-3.626, respectively,all P<0.05). The sensitivity of GCS, EEG reactivity,and relative alpha variability to predict the poor prognosis were 69.2%, 76.9%, 84.6%, respectively. The specificity were 73.3%, 80.0%, 73.3%, respectively. The consistency rates were 71.4%, 78.6%, 78.6%, respectively. The positive predictive values were 69.2%, 76.9%, 73.3%, respectively. The negative predictive values were 73.3%, 80.0%, 84.6%, respectively. More importantly, the accuracy of the relative alpha variability combined with EEG reactivity for the prediction of poor prognosis was much higher with the positive predictive value of 90.0%,the specificity of 93.3%,the sensitivity of 69.2%, the consistency rate of 82.1%,and the negative predictive values of 77.8%. Conclusions The combination of relative alpha variability and EEG reactivityis reliable to predict clinical outcome of patients with HIE.

Objective: To investigate the clinical significance of transcranial Doppler (TCD) in early diagnosis of sepsis-associated encephalopathy(SAE). Methods: Septic patients admitted to the intensive care unit(ICU) were recruited at Xiangya Hospital, Central South University from July 2015 to March 2016. Clinical data and TCD parameters during 24 hours after admission were collected. All patients were screened for delirium using the confusion assessment method for the intensive care unit (CAM-ICU) twice a day. The gold standard of the diagnosis of SAE was positive CAM-ICU evaluation. Patients were divided into SAE group and the non-SAE group. TCD data including systolic velocity (Vs), diastolic velocity (Vd), mean velocity (Vm), pulsatility index (PI) and resistant index (RI) were analyzed to determine the optimal diagnostic cut-off value. Results: A total of 43 patients were enrolled including 12 in SAE group and 31 in non-SAE group. Vm and Vd were lower in SAE group [Vm: (53.50±12.22) cm/s vs. (61.68±9.63) cm/s, P<0.05; Vd: (33.42±10.87) cm/s vs. (43.16±7.84) cm/s, P<0.01] but PI and RI were significant higher in SAE group[PI:(1.16±0.2) vs. (0.90±0.15), P<0.01;RI:(0.65±0.08) vs. (0.56±0.06), P<0.01] than in non-SAE group. The cut-off values of Vs, Vm, Vd, PI and RI for the diagnosis of SAE were 112cm/s, 55.50cm/s, 34.50cm/s, 1.16, 0.65, respectively, with the relevant sensitivities of 19.4%, 83.9%, 93.5%, 58.3%, 58.3% and the specificities of 100.0%, 50.0%, 58.3%, 96.8%, 96.8%, respectively. The diagnostic AUC of Vd, PI and RI were 0.741, 0.808 and 0.808 respectively. Conclusions The parameter changes of TCD suggest that the pathogenesis of SAE is related to cerebral hypoperfusion, TCD is a helpful method for the early diagnosis of SAE.

Objective To investigate the relationship between the serum levels of retinol-binding protein 4(RBP4),blood lipid level and glycemic control in patients with chronic schizophrenia complicated with type 2 diabetes.Methods The levels of glycated hemoglobin (HbA1c),fasting plasma glucose (FPG),cholesterol (TC),triglycerides (TG),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),apolipoproteinA1 (Apo A1),apolipoprotein B(Apo B),apolipoprotein E(Apo E),lipoprotein(a)[Lp (a)],free fatty acid(FFA),RBP4,fasting insulin (FINS) were determined in 96 patients with chronic schizo-phrenia complicated with type 2 diabetes (schizophrenia group) and 50 healthy subjects (healthy control group),the insulin resistance index (HOMA IR)was calculated.The patients were divided into HbA1c<7.0% group and HbA1c≥7.0% group according to the results of the HbA1c,and the results were statistically analyzed.Results In the HbA1c<7.0% group,the levels of HbA1c,FPG,TG,Apo B,Apo E,Lp(a),RBP4 were significantly higher than those in the healthy control group,the levels of serum Apo A1 and HDL-C was lower than those in the healthy control group (P<0.05).In the HbA1c≥7.0% group,the levels of HbA1c, Body mass index (BMI),FPG,TG,Apo E,FFA,RBP4,FINS,HOMA-IR were significantly higher than those in the HbA1c<7.0% group and the healthy control group,the levels of TC,Apo B,Lp(a)were significantly higher than the healthy control group and the levels of serum Apo A 1 and HDL-C were lower than those in the healthy control group (P<0.05).Single factor Logistic regression analysis showed that BMI,TG,Apo E, FFA,RBP4,FINS and HOMA-IR were risk factors for poor control of blood glucose in type 2 diabetes pa-tients with chronic schizophrenia.Conclusion The levels of RBP4 and blood lipid in patients with chronic schizophrenia complicated with type 2 diabetes may be related to the poor control of blood sugar level.