Objective To investigate the effect of dissociation of the human retinal pigment epithelium (RPE) cells by ficoll hypaque gradient centrifugation.Methods The primary human RPE cells and subcultured human RPE cells were dissociated with ficoll gradient centrifugation solution (d=1.077 g/ml)and the same divid ed cells as the control were dissociated with routine normal culture medium cent rifugation. The Trypan blue (0.4%) rejection staining was used, and the mouse anti-human monoclonal anti-body and fluorescein isothiocyanate (FITC) labeled rabbit anti-mouse IgG were utilized for indirect immunoreactivity for the test of human cytokeratin (CK) in active RPE cells cytoplasm. Flow cytometry assay was used to analyzed the percentages of CK positive staining RPE cells. simultaneously, the cells configuration, growth condition, the rate of clone formation, and the purifying result were observed under the fluorescent and confocal microscope.Results The survival rate and positive rate of CK of RPE cells in experimental group were higher than those in the control（P＜0.001）, but the number of the cells was reduced. The cells in the experimental group were integrated round with smooth border, symmetrical staining, homogeneous configuration and higher rate of clone formation （P＜0.001）. Conclusions RPE cells disas sociated with ficoll gradient centrifugation have the better dissociation effects. (Chin J Ocul Fundus Dis，2003，19：333-404)
For speech detection in Parkinson’s patients, we proposed a method based on time-frequency domain gradient statistics to analyze speech disorders of Parkinson’s patients. In this method, speech signal was first converted to time-frequency domain (time-frequency representation). In the process, the speech signal was divided into frames. Through calculation, each frame was Fourier transformed to obtain the energy spectrum, which was mapped to the image space for visualization. Secondly, deviations values of each energy data on time axis and frequency axis was counted. According to deviations values, the gradient statistical features were used to show the abrupt changes of energy value in different time-domains and frequency-domains. Finally, KNN classifier was applied to classify the extracted gradient statistical features. In this paper, experiments on different speech datasets of Parkinson’s patients showed that the gradient statistical features extracted in this paper had stronger clustering in classification. Compared with the classification results based on traditional features and deep learning features, the gradient statistical features extracted in this paper were better in classification accuracy, specificity and sensitivity. The experimental results show that the gradient statistical features proposed in this paper are feasible in speech classification diagnosis of Parkinson’s patients.
Pulsed magnetic field gradients generated by gradient coils are widely used in signal location in magnetic resonance imaging (MRI). However, gradient coils can also induce eddy currents in final magnetic field in the nearby conducting structures which lead to distortion and artifact in images, misguiding clinical diagnosis. We tried in our laboratory to measure the magnetic field of gradient-induced eddy current in 1.5 T superconducting magnetic resonance imaging device; and extracted key parameters including amplitude and time constant of exponential terms according to inductance-resistance series mathematical module. These parameters of both self-induced component and crossing component are useful to design digital filters to implement pulse pre-emphasize to reshape the waveform. A measure device that is a basement equipped with phantoms and receiving coils was designed and placed in the isocenter of the magnetic field. By applying testing sequence, contrast experiments were carried out in a superconducting magnet before and after eddy current compensation. Sets of one dimension signal were obtained as raw data to calculate gradient-induced eddy currents. Curve fitting by least squares method was also done to match inductance-resistance series module. The results also illustrated that pulse pre-emphasize measurement with digital filter was correct and effective in reducing eddy current effect. Pre-emphasize waveform was developed based on system function. The usefulness of pre-emphasize measurement in reducing eddy current was confirmed and the improvement was also presented. All these are valuable for reducing artifact in magnetic resonance imaging device.
Objective To quantitatively evaluate the effect of 2 types of pressures induced injury by using threedimensional (3D) reconstruction of rats loaded tibial is anterior muscle from two-dimensional (2D) image of serial histological sections. Methods Twenty female or male Sprague Dawley rats, aged 10-12 weeks and weighing 280-300 g, were randomlydivided into experimental group (n=10) and control group (n=10). The random side of tibial is anterior muscle was givenintermittent gradient (8.0-21.3 kPa) and sustained (13.3 kPa) pressure in 0.12 cm2 area in experimental group and controlgroup, respectively; the experiment was terminated and the general condition of rats was observed after 3 cycles, and a single cycle included 2 hours of compression and 30 minutes of release. The general observations of pressed skin and tibial is anterior muscle were done after 24 hours of pressure rel ief, and the tibial is anterior muscle was harvested integrally from the loaded side, then made into interval 4 μm serial sections. After HE staining, 2D images were obtained. Necrosis and injury areas were distinguished by Image Pro Plus (IPP) 6.0 software and image registration was conducted by Photoshop 8.0.1 after 2D panorama images acquired by digital microscope (× 40) and IPP mosaic software. 3D reconstruction was establ ished via data processing using Mimics 10.1 software so as to get the volume, the surface area, and 3D images of the whole piece of tibial is anterior muscle and injury areas respectively. Results All rats of 2 groups survived till experiment terminated and no skin ulcers occurred after 24 hours. Edema and indentation were observed on press side skin and tibial is anterior muscles of 2 groups, fadeless maroon area was observed in control group. A total of 994 sl ices were obtained from 20 samples of tibial is anterior muscles. 3D images suggested that injury of control group was severe, which penetrated the whole piece of tibial is anterior muscle and expandedalong the tibia bony prominence. By contrast, injury of experimental group was less, but had similar width to the contact surface of indentor. There was no significant difference in the volume and the surface area of tibial is anterior muscle between 2 groups (P gt; 0.05), while the injury volume and the injury surface area were significantly smaller in experimental group than in control group (P lt; 0.05). Conclusion 3D reconstruction is an effective method to quantitatively evaluate pathological changes inside the integrity tissue and can provide the visual basis for the mechanical property distributed in the loaded muscle. Intermittent gradient pressure can reduce deep tissue injury.
Objective To propose an innovative self-supervised learning method for vascular segmentation in computed tomography angiography (CTA) images by integrating feature reconstruction with masked autoencoding. Methods A 3D masked autoencoder-based framework was developed, where in 3D histogram of oriented gradients (HOG) was utilized for multi-scale vascular feature extraction. During pre-training, random masking was applied to local patches of CTA images, and the model was trained to jointly reconstruct original voxels and HOG features of masked regions. The pre-trained model was further fine-tuned on two annotated datasets for clinical-level vessel segmentation. Results Evaluated on two independent datasets (30 labeled CTA images each), our method achieved superior segmentation accuracy to the supervised neural network U-Net (nnU-Net) baseline, with Dice similarity coefficients of 91.2% vs. 89.7% (aorta) and 84.8% vs. 83.2% (coronary arteries). Conclusion The proposed self-supervised model significantly reduces manual annotation costs without compromising segmentation precision, showing substantial potential for enhancing clinical workflows in vascular disease management.
In developed nations, aortic stenosis (AS) is the most common valvular heart disease presentation, and its prevalence is increasing due to aging populations. Accurate diagnosis of the disease process and determination of its severity are essential in clinical decision-making. Although current guidelines recommend measuring transvalvular gradients, maximal velocity, and aortic valve area in determining the disease severity, inconsistent grading of disease severity remains a common problem in clinical practice. Recent studies suggest that patients with paradoxical low-flow and/or low-gradient, severe AS are at a more advanced stage of the disease process and have a poorer prognosis. This mode of presentation may lead to an underevaluation of symptoms and inappropriate delay of AVR. Therefore, this challenging clinical situation should be carefully assessed in particular in symptomatic patients and clinical decisions should be tailored individually.
Objective To study the effect of local appl ication of different concentrations of nerve growth factor (NGF) on fracture heal ing, and to further search for the appropriate concentration gradient of NGF to promote fracture heal ing. Methods Seventy-five adult male Sprague Dawley rats, weighing (220.0 ± 2.5) g, were made the right tibia fracture model at 1 cm distal from the tibial tubercle and randomly divided into 5 groups (groups A, B, C, D, and E, n=15). Fractures were treated with 0.3 mL normal sal ine containing different concentration of NGF (0.006 48 × 10-2, 0.032 40 × 10-2, 0.162 00 ×10-2, and 0.810 00 × 10-2 μg/g) in groups A, B, C, and D, respectively, and the same amount of normal sal ine in group E. After2, 4, and 6 weeks, the specimens were harvested from 5 rats of each group to perform the biochemical test and histological observation. Before the rats were sacrificed, the arteriovenous blood was taken from the eye-ball to test the alkal ine phosphatase (ALP) activity. Results After 2, 4, and 6 weeks, the gross observation showed that the size and hardness of bone tissue and callus tissue growth gradually increased in groups A, B, C, and D, and group D was higher than groups A, B, C, and E. The X-ray films showed that the calcified area gradually increased in groups A, B, C, and D, and group D was higher than groups A, B, C, and E. The histological observation showed that the trabecular qual ity and maturity in group D were better than those in groups A, B, C, and E. Group D was significantly higher than groups A, B, C, and E (P lt; 0.05) in the gray values of callus tissue and the calcium content of callus tissue at 4 and 6 weeks, in the wet weight of callus tissue at 2 and 4 weeks, and in the ALP content of serum at 2 weeks. The trabecula surface index of osteoblast, the trabecular volume, and the trabecular width decreased as time in the order of groups A, B, C, and D, which were higher than those of group E; group D was the highest, showing significant differences when compared with the other groups (P lt; 0.05). Conclusion The local appl ication of NGF can promote fracture heal ing in rats. The high concentration gradient of NGF (0.810 00 × 10-2 μg/g) has an obvious promotion role on fracture heal ing.
ObjectiveTo investigate the perioperative hemodynamic changes of off-pump coronary artery bypass grafting (OPCABG) patients monitored by pulse recorded analysis method (MostCare/PRAM devices) and its relationship with the prognosis.MethodsA total of 89 patients who underwent OPCABG from October 2016 to January 2017 in Beiijng Anzhen Hospital were included, including 53 males and 36 females aged 60.50±8.40 years. The hemodynamic changes were recorded. The patients were divided into two groups (a major adverse cardiovascular events group and a stable group) according to whether major adverse cardiovascular events occurred or not. The difference of hemodynamic changes between the two groups was analysed.ResultsThe mean percentage increases of stroke volume (SV) in the passive leg raising (PLR) test before opening chest and after chest closure were 23.00%±3.20% and 29.40%±3.70%, respectively. Hemodynamic data were analysed seven times, namely, anaesthesia, opening chest, heparin administration, coronary artery bypass grafting, protamine administration, thoracic closure and after operation. SV was significantly decreased during above periods, while systemic vascular resistance index (SVRI) was significantlyincreased. Cardiac circle efficiency (CCE) and maximum pressure gradient (dP/dT) were decreased after anaesthesia, and decreased to the lowest value during the procedure of bypass grafting, and then they began to increase gradually after the manipulation of bypass grafting was finished. Stroke volume variation (SVV) and pulse pressure variation (PPV) were slightly decreased during anaesthesia, then increased significantly through the whole surgery. Major adverse cardiovascular events occurred in 9 patients and 4 of them died. The basic mean values of SVRI, SVV and PPV of patients in the major adverse cardiovascular events group before opening chest were significantly higher than those of patients in the stable group. There was no significant difference in the mean values of CCE, dP/dT or SV between the two groups. There was no significant correlation between the prognosis and the mean values of SVRI, SVV, PPV, CCE, dP/dT or SV.ConclusionThe hemodynamic indexes are not stable, thus, it is necessary to monitor the perioperative hemodynamic changes of OPCABG patients timely by MostCare/PRAM device and adjust treatment measures accordingly.
ObjectiveTo investigate the value of proton magnetic resonance spectroscopy (1H-MRS), gradient dual-echo, and triple-echo sequences in the quantitative evaluation of treatment effect of fatty liver at 3.0T MR.MethodsThirty patients with fatty liver diagnosed by CT or ultrasound who admitted in Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital between August 2017 and May 2018, were enrolled and undergone gradient dual-echo, triple-echo, and 1H-MRS examination before and 3 months after treatment. The fat index (FI) and relative lipid content (RLC) were measured. Fatty liver index (FLI) was calculated from blood biochemical indicators, waist circumference, and BMI at the same time. With the reference standard of FLI, the results before and after treatment measured from MRI were analyzed.ResultsThere were significantly differences of FLI, FIdual, FItriple, and RLC before and after treatment (t=5.281, P<0.001; Z=–3.651, P<0.001; Z=–3.630, P<0.001; Z=–4.762, P<0.001), all indexes decreased after treatment. FIdual and FItriple were positively correlated with FLI before (rs=0.413, P=0.023; rs=0.396, P=0.030) and after treatment (rs=0.395, P=0.031; rs=0.519, P=0.003), the highest correlation factor was FItriple to FLI after treatment. There were no significant correlation between RLC and FLI before and after treatment (P>0.05).ConclusionsIt is feasible to quantitatively evaluate the treatment effect of fatty liver by using 1H-MRS, gradient dual-echo, and triple-echo sequences. Gradient triple-echo sequences has better accuracy, which is technically easy to implement and more suitable for clinical development.
The peak period of cardiovascular disease (CVD) is around the time of awakening in the morning, which may be related to the surge of sympathetic activity at the end of nocturnal sleep. This paper chose 140 participants as study object, 70 of which had occurred CVD events while the rest hadn’t during a two-year follow-up period. A two-layer model was proposed to investigate whether hypnopompic heart rate variability (HRV) was informative to distinguish these two types of participants. In the proposed model, the extreme gradient boosting algorithm (XGBoost) was used to construct a classifier in the first layer. By evaluating the feature importance of the classifier, those features with larger importance were fed into the second layer to construct the final classifier. Three machine learning algorithms, i.e., XGBoost, random forest and support vector machine were employed and compared in the second layer to find out which one can achieve the highest performance. The results showed that, with the analysis of hypnopompic HRV, the XGBoost+XGBoost model achieved the best performance with an accuracy of 84.3%. Compared with conventional time-domain and frequency-domain features, those features derived from nonlinear dynamic analysis were more important to the model. Especially, modified permutation entropy at scale 1 and sample entropy at scale 3 were relatively important. This study might have significance for the prevention and diagnosis of CVD, as well as for the design of CVD-risk assessment system.