ObjectiveBased on the experience of traditional quality evaluation， the quality of Atractylodis Macrocephalae Rhizoma（AMR） with different production methods such as direct seeding， transplanting after seedling raising， topping and non-topping， and difference in growth years was compared. MethodVernier caliper was used to measure the trait data of AMR in different production methods. Paraffin sections of AMR with different production methods were made by saffron solid green staining， and the microstructure was observed. The contents of water-soluble and alcohol-soluble extracts in AMR with different production methods were determined according to the 2020 edition of Chinese Pharmacopoeia. The content of water-soluble total polysaccharides in AMR with different production methods was detected by sulfuric acid-anthrone method. Fiber analyzer was used to detect the content of fiber components in AMR with different production methods. The contents of monosaccharides， oligosaccharides and some secondary metabolites in AMR with different production methods were detected by ultra performance liquid chromatography（UPLC）， and the differences of chemical components were compared by multivariate statistical analysis methods such as principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA）. ResultIn terms of traits， the 3-year-old AMR with direct seeding and without topping was close to the high-quality AMR with "phoenix-head and crane-neck， strong sweetness and clear aroma" recorded in ancient materia medica， followed by the 3-year-old AMR with topping after transplanting， while the 2-year-old AMR with topping after transplanting with high market circulation rate was generally fat and strong with mild odor. In the microscopic aspect， the arrangement of xylem vessels and fiber bundles in the 3-year-old samples formed two obvious rings. Compared with the 2-year-old samples cultivated in Bozhou and Zhejiang， the 3-year-old samples without topping after transplanting had more wood fibers. In terms of chemical composition， the contents of 70% ethanol extract， fructose， glucose， sucrose， 1-kestose， atractylenolide Ⅰ， chlorogenic acid， neochlorogenic acid， cryptochlorogenic acid and other components in 3-year-old AMR with direct seeding and without topping were significantly higher than those in the other three samples（P<0.05）. The contents of cellulose， 70% ethanol extract， sucrose， atractylenolide Ⅰ， atractylone and other components in 3-year-old AMR with topping after transplanting were significantly higher than those in the 2-year-old AMR with high market circulation rate（P<0.05）， while the contents of water-soluble extract and water-soluble total polysaccharides in 2-year-old samples with topping after transplanting were significantly higher than those in the 3-year-old AMR with topping after transplanting， direct seeding and without topping（P<0.05）. ConclusionUnder the current mainstream production mode， too much manual intervention makes AMR heavily enriched in polysaccharides and increased the yield， but the accumulation of sweet substances， fragrant substances and fiber substances is insufficient， which affects its quality. The current quality standard of AMR has some shortcomings in guiding the high quality production of it， it is suggested to revise the quality standard of AMR， supplement the quantitative analysis of secondary metabolites， and strengthen the production of imitation wild AMR.

Objective To develop a multi-modal deep learning method for automatic classification of immune-mediated glomerular diseases based on images of optical microscopy(OM),immunofluorescence microscopy(IM),and transmission electron microscopy(TEM).Methods We retrospectively collected the pathological images from 273 patients and constructed a multi-modal multi-instance model for classification of 3 immune-mediated glomerular diseases,namely immunoglobulin A nephropathy(IgAN),membranous nephropathy(MN),and lupus nephritis(LN).This model adopts an instance-level multi-instance learning(I-MIL)method to select the TEM images for multi-modal feature fusion with the OM images and IM images of the same patient.By comparing this model with unimodal and bimodal models,we explored different combinations of the 3 modalities and the optimal methods for modal feature fusion.Results The multi-modal multi-instance model combining OM,IM,and TEM images had a disease classification accuracy of(88.34±2.12)%,superior to that of the optimal unimodal model[(87.08±4.25)%]and that of the optimal bimodal model[(87.92±3.06)%].Conclusion This multi-modal multi-instance model based on OM,IM,and TEM images can achieve automatic classification of immune-mediated glomerular diseases with a good classification accuracy.

Objective To develop a multi-modal deep learning method for automatic classification of immune-mediated glomerular diseases based on images of optical microscopy(OM),immunofluorescence microscopy(IM),and transmission electron microscopy(TEM).Methods We retrospectively collected the pathological images from 273 patients and constructed a multi-modal multi-instance model for classification of 3 immune-mediated glomerular diseases,namely immunoglobulin A nephropathy(IgAN),membranous nephropathy(MN),and lupus nephritis(LN).This model adopts an instance-level multi-instance learning(I-MIL)method to select the TEM images for multi-modal feature fusion with the OM images and IM images of the same patient.By comparing this model with unimodal and bimodal models,we explored different combinations of the 3 modalities and the optimal methods for modal feature fusion.Results The multi-modal multi-instance model combining OM,IM,and TEM images had a disease classification accuracy of(88.34±2.12)%,superior to that of the optimal unimodal model[(87.08±4.25)%]and that of the optimal bimodal model[(87.92±3.06)%].Conclusion This multi-modal multi-instance model based on OM,IM,and TEM images can achieve automatic classification of immune-mediated glomerular diseases with a good classification accuracy.

OBJECTIVE: We propose a novel region- level self-supervised contrastive learning method USRegCon (ultrastructural region contrast) based on the semantic similarity of ultrastructures to improve the performance of the model for glomerular ultrastructure segmentation on electron microscope images. METHODS: USRegCon used a large amount of unlabeled data for pre- training of the model in 3 steps: (1) The model encoded and decoded the ultrastructural information in the image and adaptively divided the image into multiple regions based on the semantic similarity of the ultrastructures; (2) Based on the divided regions, the first-order grayscale region representations and deep semantic region representations of each region were extracted by region pooling operation; (3) For the first-order grayscale region representations, a grayscale loss function was proposed to minimize the grayscale difference within regions and maximize the difference between regions. For deep semantic region representations, a semantic loss function was introduced to maximize the similarity of positive region pairs and the difference of negative region pairs in the representation space. These two loss functions were jointly used for pre-training of the model. RESULTS: In the segmentation task for 3 ultrastructures of the glomerular filtration barrier based on the private dataset GlomEM, USRegCon achieved promising segmentation results for basement membrane, endothelial cells, and podocytes, with Dice coefficients of (85.69 ± 0.13)%, (74.59 ± 0.13)%, and (78.57 ± 0.16)%, respectively, demonstrating a good performance of the model superior to many existing image-level, pixel-level, and region-level self-supervised contrastive learning methods and close to the fully- supervised pre-training method based on the large- scale labeled dataset ImageNet. CONCLUSION USRegCon facilitates the model to learn beneficial region representations from large amounts of unlabeled data to overcome the scarcity of labeled data and improves the deep model performance for glomerular ultrastructure recognition and boundary segmentation.
Humans ; Electrons ; Endothelial Cells ; Learning ; Podocytes ; Kidney Diseases

Objective    To explore the feasibility of goat tricuspid regurgitation (TR) model by one chordae tendineae cutter via right anterior-lateral minimal incision. Methods    TR model was established in 6 goats with a self-made tricuspid valve chordae tendineae cutter. The goats were placed in a left lateral position and procedure was performed via a right anterior-lateral minimal thoracotomy in the fourth intercostal. Under the guidance of transesophageal ultrasound and digital subtraction angiography, the chordae tendineae of anterior leaflet was cut until moderate to severe regurgitation was confirmed. Echocardiography and laboratory examinations were performed preoperatively, immediately and 3 months after surgery. Additionally, all goats were sacrificed to clarify pathological evaluation. Results    TR was successfully established in 6 goats. The right atrium pressure increased significantly immediately after surgery (P<0.05). During a follow-up of 3 months, the progression of TR was aggravated (P<0.05), and the annular diameter increased from 2.15±0.23 cm to 2.65±0.20 cm. Overall, there was no statistically significant change in transvalvular gradient and velocity between preoperation and postoperation. Laboratory test results showed no abnormalities between preoperation and postoperation. Autopsy evaluation demonstrated obvioue chordae tendineae transection of the anterior leaflet. Conclusion    It is feasible to establish TR model via a right minimal anterior lateral thoracotomy in the fourth intercostal space. This novel TR goats model will allow investigation of transcatheter interventional device and serve as a chronic model in the future.

Recently, several transcatheter devices for aortic valve replacement and mitral valve repair have been used in clinic, and researchers have designed a variety of tricuspid valve (TV) intervention devices. We reviewed the current status of transcatheter TV intervention, and focused on the structures of these devices and the early results of clinical trials. Undoubtedly, transcatheter intervention for TV is promising, innovational and safe for patients with severe TV regurgitation.

Objective To investigate the potential of polylactic acid glycolic acid copolymer (PLGA) composite ordered multi tunnel collagen scaffold in fabricating a biomimetic artificial nerve graft to repair the sciatic nerve defects in rats.Methods The ordered multi tunnel collagen scaffold was prepared by vacuum freeze-drying and directional drawing method to simulate the epineurium;the outer conduit was prepared by PLGA to simulate the epineurium;and then,the ordered multi tunnel collagen scaffolds were loaded in the PLGA conduit (5∶1) under a stereomicroscope to develop a novel biomimetic artificial nerve.Sixty-four rats were randomly divided into four groups:artificial nerve group,PLGA group,peripheral nerve group,and non-graft control group (n=16);rats in the artificial nerve group,PLGA group,and peripheral nerve group were repaired with artificial nerve graft,hollow PLGA conduit and allogeneic sciatic nerve to bridge the sciatic nerve defect,while the sciatic nerve with the gap in rats of the control group was without any grafting.After 11 weeks of operation,the hind limbs of rats in each group were detected by behavioral test,electrophysiological examination and Fluoro-Gold retrograde tracing method.The changes of muscle tissues (gastrocnemius) were observed by hematoxylin staining and TMR-α-BTX staining,and the regenerated axons were observed by immunohistochemical staining with NF200 and the regenerative spinal anterior horn motor neurons were observed by Nissl fluorescence staining 12 weeks after operation.Results After 11 weeks of operation,the recoveries of the motor functions (the distance between the injured hindlimb and forelimb,the rotation angle of the injured foot) in the peripheral nerve group,artificial nerve group,PLGA group and control group were significantly deteriorated in turn,and the differences were statistically significant (P＜0.05).Electrophysiological examination showed that the recovery effect of peripheral nerve group was the best in both latency and amplitude of the compound muscle action potential,followed by artificial nerve group.The latency of PLGA group was the longest and the amplitude of compound action potential was the smallest;significant differences were noted between each two groups (P2＜0.05).At 12 weeks after operation,the wet weight ratio of muscle fibers,area of muscle fibers and neuromuscular junction area were significantly different between each two groups (P＜0.05);the degree of gastrocnemius atrophy in the artificial nerve group was significantly improved than that in the PLGA group,but not yet reached the level of peripheral nerve group.NF200 immunohistochemical staining showed that a large number of NF200-positive axons were seen in the grafts of the artificial nerve group,but the number was slightly smaller than that of the peripheral nerve group;the number of regenerated axons in the PLGA group was the smaller and mainly distributed near the proximal side.In the PLGA group,only (19.33 ±6.73)％ regenerated spinal anterior horn motor neurons were labeled with Fluoro-Gold,while the positive rates of Fluoro-Gold in the artificial nerve group and peripheral nerve group were (42.67±7.45)％ and (50.13±4.33)％;the differences between each two groups were statistically significant (P＜0.05).Conclusion The biomimetic artificial nerve made of PLGA conduit and ordered multi tunnel collagen scaffold can efficiently reconstruct the defected peripheral nerve with guiding axonal regeneration and promoting functional restoration in rats;however,its effect is poor than peripheral nerve grafting.

Objective To observe the effect of Sanqi Oral Liquid on the podocytes and the expression of slit diaphragm-associated molecules (including Nephrin, Podocin and CD2AP proteins) of diabetes mellitus (DM) rats after unilateral nephrectomy, so as to explore its mechanism for protecting renal function. Methods SD male rats were randomized into sham operation group, model group and Chinese medicine group. The experimental diabetes mellitus ( DM) model was given unilateral nephrectomy and intraperitoneal injection of 35 mg/kg of streptozocin (STZ). And then the model rats were given intragastric administration of Sanqi Oral Liquid (2.5 g· kg-1·d-1) and the same volume distilled water respectively for 8 weeks. After treatment, the density of podocytes and the podocytic foot process width of different groups were measured. Immunohistochemistry was used to observe the changes of the expression of Nephrin, Podocin and CD2AP in renal tissues of different groups. Results After treatment with Sanqi Oral Liquid, the density of podocytes was increased, the foot process lesion was relieved, and the expression levels of Nephrin, Podocin and CD2AP proteins were increased (P<0.05 or P<0.01 as compared with those in the model group). Conclusion The protective mechanism of Sanqi Oral Liquid for renal function of unilateral nephrectomy DM rats is possibly related with the alleviation of podocyte injury and with the regulation of the expression of Nephrin, Podocin and CD2AP proteins in podocytes.

OBJECTIVETo explore the effects of hyperbaric oxygen preconditioning (HBOP) on human stress responses during acute exposure to high altitude and the possible mechanism.

METHODSEight male subjects were treated with HBOP for 3, 5, and 7 days, followed by acute exposure to hypoxia simulating an altitude of 4,000 m. Subjects at rest were divided into sea-level control group, simulated high-altitude group, and 5-day HBOP intervention group, while subjects after physical load were divided into sea-level control group, simulated high-altitude group, 3-day HBOP intervention group, and 7-day HBOP intervention group. The physical load test was performed for each subject before and after HBOP, and the plasma levels of dopamine (DA), epinephrine (E), norepinephrine (NE), and adrenocorticotropic hormone (ACTH) were determined before and after exercise. The physical load test was performed by stepping up on to a 30 cm-high stepping stool at a rate of 25/min for 5 minutes, which was a type of moderate physical exercise. The stepping rate and timing were controlled by a metronome.

RESULTSThe levels of DA, E, NE, and ACTH at rest and after physical load were significantly higher in subjects acutely exposed to high altitude than in the sea-level control groups (all P<0.05). Moreover, the levels of DA, E, NE, and ACTH at rest were significantly higher after acute exposure to high altitude in the 5-day HBOP intervention group than in the simulated high-altitude group (all P<0.01). Except for the ACTH level in the 3-day HBOP intervention group, the levels of DA, E, NE, and ACTH after physical load were significantly higher after acute exposure to high altitude in the 3-day and 7-day HBOP intervention groups than in the simulated high-altitude group (all P<0.01).

CONCLUSIONHBOP can elevate the plasma expression of DA, E, NE, and ACTH, and then speed up the establishment of a new balance of homeostasis to adapt to the acute hypoxia at high altitude.

Adrenocorticotropic Hormone ; blood ; Altitude ; Dopamine ; blood ; Epinephrine ; blood ; Exercise ; Homeostasis ; Humans ; Hyperbaric Oxygenation ; Hypoxia ; blood ; Male ; Norepinephrine ; blood ; Rest ; Stress, Physiological

We proposed a new stitching method based on sift features to obtain an enlarged view of transmission electron microscopic (TEM) images with a high resolution. The sift features were extracted from the images, which were then combined with fitted polynomial correction field to correct the images, followed by image alignment based on the sift features. The image seams at the junction were finally removed by Poisson image editing to achieve seamless stitching, which was validated on 60 local glomerular TEM images with an image alignment error of 62.5 to 187.5 nm. Compared with 3 other stitching methods, the proposed method could effectively reduce image deformation and avoid artifacts to facilitate renal biopsy pathological diagnosis.
Algorithms ; Artifacts ; Humans ; Image Processing, Computer-Assisted ; methods ; Kidney Glomerulus ; ultrastructure ; Microscopy, Electron, Transmission ; methods