Search

In order to better understand the particle breakage mechanics and characterization methods of calcareous sand, the particle breakage characteristics of calcareous sand in one-dimensional compression tests, isotropic compression tests, triaxial shear tests, and ring shear tests are reviewed and analyzed. The results show that the mechanism of shear crushing is essentially different from compression crushing. Compared with one-dimensional compression, it is easier to break calcareous sand under triaxial shear. In the shearing process, the particle breakage of calcareous sand will not continue indefinitely. The gradation tends to be stable and controlled by confining pressure and shear strain. The characteristic particle size measurement method for particle breakage is simple and has limitations. The description method using the particle size distribution curve is more comprehensive, but it is impossible to compare the crushing degree of particle with different particle sizes, and a suitable measurement method needs to be proposed.

Upper limb motor dysfunction after stroke is a serious threat to the living quality of patients and their families. Recovery of upper limb motor function after stroke largely relies on the activation and remodeling of neural circuits. rTMS (repetitive transcranial magnetic stimulation) has been proved to promote the reconstruction of neural synapses and neural circuits. However, there are still a large number of patients who cannot fully recover and leave behind varying degrees of dysfunction. Considering the systemic pathology after stroke, in addition to focal brain injury, stroke can also cause extensive dysfunction of peripheral organs. The rehabilitation strategy for stroke should combine the treatment of primary brain lesions with the intervention of secondary systemic damage. The aim of this trial is to verify the efficacy of rTMS synergize with Tui Na (Chinese Massage) on upper limb motor function after ischemic stroke, and to explore the mechanism of activation and remodeling of sensorimotor neural circuits with functional near-infrared spectroscopy. Ninety patients will be randomly assigned to either rTMS + Tui Na + conventional rehabilitation group (the experimental group) or rTMS + conventional rehabilitation group (the control group) in 1:1 ratio. Intervention is conducted five sessions a week, with a total of twenty sessions. The primary outcome is Fugl-Meyer Assessment, and the secondary outcomes include Muscle Strength, Modified Ashworth Assessment, Modified Barthel Index Assessment, motor evoked potentials and functional near-infrared spectroscopy. There are four time points for the evaluation, including baseline, 2 weeks and 4 weeks after the start of treatment, and 4 weeks after the end of treatment. This study is a randomized controlled trial. This study was approved by Institutional Ethics Committee of Shanghai Third Rehabilitation Hospital Affiliated to Shanghai University of Traditional Chinese Medicine (approval No. SH3RH-2021-EC-012) on December, 16th, 2021. The protocol was registered with Chinese Clinical Trial Registry (ChiCTR2200056266), on February 3th, 2022. Patient recruitment was initiated on February 10th, 2022, and the study will be continued until December 2023.

Multi-target neural circuit-magnetic stimulation has been clinically shown to improve rehabilitation of lower limb motor function after spinal cord injury. However, the precise underlying mechanism remains unclear. In this study, we performed double-target neural circuit-magnetic stimulation on the left motor cortex and bilateral L5 nerve root for 3 successive weeks in a rat model of incomplete spinal cord injury caused by compression at T10. Results showed that in the injured spinal cord, the expression of the astrocyte marker glial fibrillary acidic protein and inflammatory factors interleukin 1β, interleukin-6, and tumor necrosis factor-α had decreased, whereas that of neuronal survival marker microtubule-associated protein 2 and synaptic plasticity markers postsynaptic densification protein 95 and synaptophysin protein had increased. Additionally, neural signaling of the descending corticospinal tract was markedly improved and rat locomotor function recovered significantly. These findings suggest that double-target neural circuit-magnetic stimulation improves rat motor function by attenuating astrocyte activation, thus providing a theoretical basis for application of double-target neural circuit-magnetic stimulation in the clinical treatment of spinal cord injury.

Objective The aim of this study is to assess the neural interconnection between the acupoint “Chengshān (承山BL57)” and sciatic nerve from the peripheral nervous system (PNS) to the central nervous system (CNS). Methods SD rats were systematically examined from the hip to the hind limb along the route of sciatic nerve with regional anatomical technique, and further traced from the BL57 and the trunk of sciatic nerve by the neural tracing technique with cholera toxin subunit B (CTB) and CTB conjugated Alexa Fluor 488 and 594 (CTB-AF488/594). Results After regional anatomy, it was found that the sciatic nerve ran down from the hip to the hind limb and sequentially sent out the tibial and sural branches respectively to the deep and superficial layers of BL57. The CTB labeling associated with both BL57 and sciatic nerve distributed in the same spinal segments and definite regions, including the sensory neurons in lumber 3 (L3) -lumber 6 (L6) dorsal root ganglia (DRGs), transganglionic axons mainly in the L3-L6 spinal dorsal horn, Clarke's nucleus, and gracile nucleus, as well as motor neurons mainly in the L3-L6 spinal ventral horn. However, the amount of neural labeling was significantly less in the cases of BL57 than that of sciatic nerve. Conclusions These results indicate that there is distinct sensory and motor interconnection between the BL57 and the sciatic nerve, which may potentially serve for the active role of the BL57 playing in meliorating the disorders of sciatic nerve.

Objective: To visualize and compare the sensory and autonomic innervation of the local tissues at the sites of different traditional acupuncture points in the rat forehead and face by histochemical examination. Methods: GB14 ( Yangbai), ST2 ( Sibai) and ST6 ( Jiache) were selected as the representative traditional acupuncture points in this study, and the local tissues at these sites were dissected in rats after perfusion followed by double or triple fluorescent histochemical staining. Here, calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH) and vesicular acetylcholine transporter (VAChT) were used to label the sensory, sympathetic and parasympathetic nerve fibers, respectively. Results: The CGRP+ sensory, TH+ sympathetic and VAChT+ parasympathetic nerve fibers were simultaneously demonstrated in the local tissues at GB14, ST2 and ST6. Although the three kinds of nerve fibers ran in parallel or intermingled with each other, by the analysis from the view of three-dimensional reconstruction, it was clear that each of them distributed in an independent pattern to their corresponding target tissues including the blood vessels, hair follicles, arrector pili and subcutaneous muscles, as well as sebaceous glands. Conclusion: Our study demonstrated the sensory and autonomic innervation of the local tissues at GB14, ST2 and ST6, providing neurochemical evidence indicating that the CGRP+ sensory, TH+ sympathetic and VAChT+ parasympathetic nerve fibers form a neural network at these point locations that may respond to acupuncture stimulation.

The neurovascular structures in the cranial dura mater have been studied with various histological techniques in the past years. In order to obtain a proper approach to reveal the detailed structures, different labeling methods for the cranial vessels and nerve fibers were tested in this study. Firstly, the labeling characteristics of phalloidin, alpha smooth muscle actin (α-SMA), and CD31 were compared in rat whole-mount cranial dura mater by using fluorescent immunohistochemistry or histochemistry. Secondly, according to their properties, phalloidin and α-SMA were selected to combine with calcitonin gene-related peptide (CGRP) to further demonstrate the cranial neurovascular structure. By these approaches, a three-dimensional map of blood vessels and nerve fibers within the whole-mount rat cranial dura mater was obtained. The results showed that phalloidin, α-SMA, and CD31 were preferably expressed in the wall of cranial vessels, corresponding to the arteriors, venules, and capillaries, respectively. Additionally, CGRP + nerve fibers were clearly demonstrated together with phalloidin + or α-SMA + vessels, forming a delicate neurovascular network in the cranial dura mater. The thick nerve bundles ran closely to the phalloidin + or α-SMA + vessels in parallel pattern, while the thin nerve fibers branched off from the bundles tending to surround the phalloidin + arterioles rather than α-SMA + venules. These findings suggest that phalloidin could be an appropriate biochemical maker to be effectively used together with CGRP for experiments examining the detailed spatial correlation of cranial blood vessels and nerve fibers in a three-dimensional view, which may provide clues for understanding the underlying mechanisms of cranial neurovascular disorders.

Objective The aim of this study was to compare the microstructure of blood vessels and nerve fibers between different acupoints in the rat skin from the perspective of histology. Methods The skin tissue at “Qūze” (曲泽 PC3)” “Daling (大陵 PC7)” and “Zhōngchōng (中冲 PC9)” regions was taken from the rat forelimb along the pericardium meridian, and cut sagittally with a freezing microtome. After that, the skin sections were stained by fluorescent histochemistry and immunohistochemistry with phalloidin and calcitonin gene-related peptide (CGRP) to reveal the microstructure of blood vessels and nerve fibers respectively, and then examined under a laser scanning confocal microscope. Results The microstructure of blood vessels and nerve fibers was clearly labeled with phalloidin and CGRP, respectively. The blood vessels and nerve fibers were observed in the dermis of “PC3” “PC7” and “PC9” regions, where the bundle of nerve fibers distribute in parallel to the blood vessels, and sent out thin branches to surround the wall of the blood vessels. In contrast, the density of blood vessels and nerve fibers in “PC3” “PC7” and “PC9” were presented orderly in an increased tendency. Conclusion The results of present study demonstrate the differences of the neurovascular microstructure among “PC3” “PC7” and “PC9” in the rat, providing a histological view to insight into the specificity of different acupoints.

The geological conditions of the Bortala River basin are complex, and the transformation of surface water and groundwater is frequent. It is great significance to study the hydrochemical characteristics and transformation relationship of surface water and groundwater for the reasonable development and allocation of water resources in the basin. Based on 15 surface water samples and 39 groundwater samples collected in April and May 2021, the APCS/MLR model and the combination of stable hydrogen and oxygen isotopes with hydrochemistry were used in this study to analyze the hydrochemical types and the distribution characteristics of hydrogen and oxygen isotopes, the source contribution of chemical components, and the transformation relationship between surface water and groundwater. The results showed that the surface water was mainly HCO

BackgroundUpper-extremity hemiplegia after stroke remains a significant clinical problem. The supplementary motor area (SMA) is vital to the motor recovery outcomes of chronic stroke patients. Therefore, rebuilding the descending motor tract from the SMA to the paralyzed limb is a potential approach to restoring arm motor function after stroke. Paired associative stimulation (PAS), which is based on Hebbian theory, is a potential method for reconstructing the connections in the impaired motor neural circuits. The study described in this protocol aims to assess the effects of cortico–peripheral Hebbian-type stimulation (HTS), involving PAS, for neural circuit reconstruction to rescue the paralyzed arm after stroke.MethodsThe study is a 4-month double-blind randomized sham-controlled clinical trial. We will recruit 90 post-stroke individuals with mild to moderate upper limb paralysis. Based on a 1:1 ratio, the participants will be randomly assigned to the HTS and sham groups. Each participant will undergo 5-week HTS or sham stimulation. Assessments will be conducted at baseline, immediately after the 5-week treatment, and at a 3-month follow-up. The primary outcome will be the Wolf Motor Function Test (WMFT). The secondary outcomes will be Fugl-Meyer Assessment for Upper Extremity (FMA-UE), Functional Independence Measure (FIM), and functional near-infrared spectroscopy (fNIRS) parameters. The adverse events will be recorded throughout the study.DiscussionUpper-limb paralysis in stroke patients is due to neural circuit disruption, so the reconstruction of effective motor circuits is a promising treatment approach. Based on its anatomical structure and function, the SMA is thought to compensate for motor dysfunction after focal brain injury at the cortical level. Our well-designed randomized controlled trial will allow us to analyze the clinical efficacy of this novel Hebbian theory-based neuromodulation strategy regarding promoting the connection between the cortex and peripheral limb. The results may have significance for the development and implementation of effective neurorehabilitation treatments.Clinical Trial Registration[www.ClinicalTrials.gov], identifier [ChiCTR2000039949].

Characterized by high morbidity, mortality, and disability, stroke usually causes symptoms of cerebral hypoxia due to a sudden blockage or rupture of brain vessels, and it seriously threatens human life and health. Rehabilitation is the essential treatment for post-stroke patients suffering from functional impairments, through which hemiparesis, aphasia, dysphagia, unilateral neglect, depression, and cognitive dysfunction can be restored to various degrees. Noninvasive brain stimulation (NIBS) is a popular neuromodulatory technology of rehabilitation focusing on the local cerebral cortex, which can improve clinical functions by regulating the excitability of corresponding neurons. Increasing evidence has been obtained from the clinical application of NIBS, especially repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (tDCS). However, without a standardized protocol, existing studies on NIBS show a wide variation in terms of stimulation site, frequency, intensity, dosage, and other parameters. Its application for neurorehabilitation in post-stroke patients is still limited. With advances in neuronavigation technologies, functional near-infrared spectroscopy, and functional MRI, specific brain regions can be precisely located for stimulation. On the basis of our further understanding on neural circuits, neuromodulation in post-stroke rehabilitation has also evolved from single-target stimulation to co-stimulation of two or more targets, even circuits and the network. The present study aims to review the findings of current research, discuss future directions of NIBS application, and finally promote the use of NIBS in post-stroke rehabilitation.

Modified constraint-induced movement therapy (mCIMT) has shown beneficial effects on motor function improvement after brain injury, but the exact mechanism remains unclear. In this study, amplitude of low frequency fluctuation (ALFF) metrics measured by resting-state functional magnetic resonance imaging was obtained to investigate the efficacy and mechanism of mCIMT in a control cortical impact (CCI) rat model simulating traumatic brain injury. At 3 days after control cortical impact model establishment, we found that the mean ALFF (mALFF) signals were decreased in the left motor cortex, somatosensory cortex, insula cortex and the right motor cortex, and were increased in the right corpus callosum. After 3 weeks of an 8-hour daily mCIMT treatment, the mALFF values were significantly increased in the bilateral hemispheres compared with those at 3 days postoperatively. The mALFF signal values of left corpus callosum, left somatosensory cortex, right medial prefrontal cortex, right motor cortex, left postero dorsal hippocampus, left motor cortex, right corpus callosum, and right somatosensory cortex were increased in the mCIMT group compared with the control cortical impact group. Finally, we identified brain regions with significantly decreased mALFF values at 3 days postoperatively. Pearson correlation coefficients with the right forelimb sliding score indicated that the improvement in motor function of the affected upper limb was associated with an increase in mALFF values in these brain regions. Our findings suggest that functional cortical plasticity changes after brain injury, and that mCIMT is an effective method to improve affected upper limb motor function by promoting bilateral hemispheric cortical remodeling. mALFF values correlate with behavioral changes and can potentially be used as biomarkers to assess dynamic cortical plasticity after traumatic brain injury.

The schematic diagram is an indispensable part of research article in life science，which can effectively and intuitively show the specific content of the article with simple composition. As an intuitive expression of the interdisciplinary subject at the early stage, the schematic diagram in the literature has gained an all-round improvement in the quality along with the deve-lopment of computer technology and modern drawing tools. In contrast，the level of the schematic diagram appears to be lagged behind in the field of acupuncture research papers. In order to improve the drawing level of this field, we took the Five Shu-points of Hand and Foot Jueyin Meridian as an example, and drew some vector diagrams of their anatomical structures including the body surface, skeleton, nerves, arteries and veins by using the Adobe Illustrator image software, through which we sum up a simple and easy-to-learn process including application skills and key points needing attention. We hope these methods can play a role in the acupuncture research in the future.示意图是现代生命科学研究中不可缺少的重要组成部分，它以简洁的构图方式有效且清晰地展示出所表达的具体内容。作为较早的交叉学科发展的直观体现，随着计算机技术的进步和现代绘图工具的不断涌现，在生命科学文献中所看到示意图的质量已经得到了全面提升。相比之下，示意图的水准在针灸学领域显得有所滞后。为了有效提升针灸学领域示意图的绘制水平，本研究以人体手足厥阴经五输穴相关解剖学结构为例，运用Adobe Illustrator绘图软件从体表、骨骼、神经和动静脉几个方面尝试绘制矢量图。通过示意图的绘制，我们总结了简单易学的绘制流程、应用技巧及注意事项等，期待这些方法在今后针灸研究工作中可以发挥更大的作用。.

Objective The innervations in Yīnlingquan (阴陵泉 SP9) and Yanglingquan (阳陵泉 GB34) were investigated to explore the neuroanatomical interconnection of the complementary acupoints in the rat hindlimb. Methods A dual neural tracing technique was used, six male Sprague-Dawley rats were included in experiment. Under anesthesia with isoflurane, cholera toxin subunit B conjugated with Alexa Fluor 488 and 594 (CTB-AF488/594) were separately injected into SP9 and GB34 in the same rat. Three days later, the neural labeling was examined in the dorsal root ganglion (DRG), spinal cord, and sympathetic chain at lumbar (L) spinal segments under a laser scanning confocal microscope. Results All neural labeling were present on the ipsilateral side of tracer injection, including sensory, motor and sympathetic postganglionic neurons. The sensory neurons associated with SP9 and GB34 were distributed in the DRG from L2 to L6 and concentrated in L3-L4 and L4-L5 respectively, while most of the double labeled sensory neurons were presented in L4. On the other hand, the labeled motor neurons for SP9 and GB34 were observed in the dorsolateral part of spinal ventral horn at L2-L4 and L3-L5 respectively, and some of them were double labeled at L4. In addition, the sympathetic postganglionic neurons related to SP9 and GB34 were found in the lumbar sympathetic chain respectively. Conclusion The similarities of the sensory, motor, and sympathetic innervation of SP9 and GB34 in the rat provide the neuroanatomical evidence to understand the synergetic effect that arise from the stimulation at the complementary acupoints.

Investigating the soil-water characteristics of calcareous soil has a great significance for preventing geological disasters on island-reefs as well as maintaining the foundation stability of hydraulic-filled island-reefs. In this study, calcareous silty sands with different fines contents and dry densities were studied to reveal their effects on the soil-water characteristics of calcareous soil on hydraulic-filled island-reefs. The soil-water characteristic curve (SWCC) of the calcareous silty sand was measured using a pressure plate apparatus. Taking into account the porous meso-structure, the effects of fines content and dry density on the SWCC of calcareous silty sand were analyzed, and the applicability of existing SWCC models to calcareous silty sand was verified. A SWCC model suitable for assessing soil-water characteristic of calcareous silty sand was proposed. Results of this study provide some reference for quantifying the water-holding capacity of calcareous silty sand.

The aim of this study was to examine the distribution and origin of the calcitonin gene-related peptide (CGRP)-immunoreactive sensory nerve fibers of the cranial dura mater using immunofluorescence, three-dimensional (3D) reconstruction and retrograde tracing technique. Here, the nerve fibers and blood vessels were stained using immunofluorescence and histochemistry techniques with CGRP and fluorescent phalloidin, respectively. The spatial correlation of dural CGRP-immuoreactive nerve fibers and blood vessels were demonstrated by 3D reconstruction. Meanwhile, the origin of the CGRP-immunoreactive nerve fibers were detected by neural tracing technique with fluorogold (FG) from the area around middle meningeal artery (MMA) in the cranial dura mater to the trigeminal ganglion (TG) and cervical (C) dorsal root ganglia (DRGs). In addition, the chemical characteristics of FG-labeled neurons in the TG and DRGs were also examined together with CGRP using double immunofluorescences. Taking advantage of the transparent whole-mount sample and 3D reconstruction, it was shown that CGRP-immunoreactive nerve fibers and phalloidin-labeled arterioles run together or separately forming a dural neurovascular network in a 3D view, while the FG-labeled neurons were found in the ophthalmic, maxillary, and mandibular branches of TG, as well as the C2-3 DRGs ipsilateral to the side of tracer application in which some of FG-labeled neurons presented with CGRP-immunoreactive expression. With these approaches, we demonstrated the distributional characteristics of CGRP-immunoreactive nerve fibers around the blood vessels in the cranial dura mater, as well as the origin of these nerve fibers from TG and DRGs. From the perspective of methodology, it may provide a valuable reference for understanding the complicated neurovascular structure of the cranial dura mater under the physiological or pathological condition.

Encouraging results have been reported for the use of transcranial magnetic stimulation-based nerve stimulation in studies of the mechanisms of neurological regulation, nerve injury repair, and nerve localization. However, to date, there are only a few reviews on the use of transcranial magnetic stimulation for diabetic neuropathy. Patients with diabetic neuropathy vary in disease progression and show neuropathy in the early stage of the disease with mild symptoms, making it difficult to screen and identify. In the later stage of the disease, irreversible neurological damage occurs, resulting in treatment difficulties. In this review, we summarize the current state of diabetic neuropathy research and the prospects for the application of transcranial magnetic stimulation in diabetic neuropathy. We review significant studies on the beneficial effects of transcranial magnetic stimulation in diabetic neuropathy treatment, based on the outcomes of its use to treat neurodegeneration, pain, blood flow change, autonomic nervous disorders, vascular endothelial injury, and depression. Collectively, the studies suggest that transcranial magnetic stimulation can produce excitatory/inhibitory stimulation of the cerebral cortex or local areas, promote the remodeling of the nervous system, and that it has good application prospects for the localization of the injury, neuroprotection, and the promotion of nerve regeneration. Therefore, transcranial magnetic stimulation is useful for the screening and early treatment of diabetic neuropathy. Transcranial magnetic stimulation can also alleviate pain symptoms by changing the cortical threshold and inhibiting the conduction of sensory information in the thalamo-spinal pathway, and therefore it has therapeutic potential for the treatment of pain and pain-related depressive symptoms in patients with diabetic neuropathy. Additionally, based on the effect of transcranial magnetic stimulation on local blood flow and its ability to change heart rate and urine protein content, transcranial magnetic stimulation has potential in the treatment of autonomic nerve dysfunction and vascular injury in diabetic neuropathy. Furthermore, oxidative stress and the inflammatory response are involved in the process of diabetic neuropathy, and transcranial magnetic stimulation can reduce oxidative damage. The pathological mechanisms of diabetic neuropathy should be further studied in combination with transcranial magnetic stimulation technology.

To study the effect of bone marrow mesenchyml stem cell (BMSC) exosomes (Exo) on the proliferation and migration of brain microvascular endothelial cells in rats.BMSCs were extracted from rats and identified. The BMSCs were co-cultured with bEnd.3 cells in Transwell chamber for 24 h (BMSCs group). Extracted and identified the BMSCs exosomes (BMSC-Exo). Observed and qualitatively evaluated the cells' abilities on swallowing the BMSC-Exo under a fluorescence microscope. The optimal work concentration of BMSC-Exo was selected by detecting the cell vitality under different BMSC-Exo concentrations by CCK8 method. bEnd.3 cells were co-cultured with BMSC-Exo for 24 h (BMSC-Exo group). bEnd.3 cells cultured alone was set as control group. The proliferation and migration of bEnd.3 cells in the three groups were respectively detected by EDU and cell scratching experiment after 24 h of culture.Flow cytometry showed that P3 BMSCs were CD90 and CD29 positive and CD45 negative, with osteogenic differentiation and adipogenesis differentiation, indicating the extracted BMSCs high purity. The BMSC-Exo under transmission electron microscopy was round-shaped with a diameter of about 100 nm; NTA analysis found the diameter distribution of BMSC-Exo ranged from 50 to 600 nm, with a peak size of 150 nm. Immunofluorescence showed that the endothelial cells could swallow BMSC-Exo. CCK8 showed that supplement of 20 μg/mL BMSC-Exo had the best effect on cell proliferation. EDU results showed that BMSCs group and BMSC-Exo group could promote the proliferation of bEnd.3 cells compared with the control group (BMSC-Exo can replace BMSCs in effectively promoting the proliferation and migration of cerebral microvascular endothelial cells, which provide a new potential treatment for angiogenesis after stroke.

Motor function injuries mostly block or interfere with motor nerve conduction pathways. Theinnovative multi-target magnetic stimulation (MS) technology stimulates the cortex and nerve roots to activate the human central and peripheral nervous system, and has achieved preliminary results in scientificand clinical exploration. The mechanismsof these researches involve the activation of neurons, nerve conduction, the regulation of the microenvironment of the injured area, and gene regulation. However, how to make the multi-target MS technology achieve more accurate and effective nerve repair in rehabilitation is a bottleneck problem, which may need to break through neural circuit reconstruction and task-oriented training. This topic focuses on Nerve Regeneration Modulation and Rehabilitation, aiming to apply the concepts of nerve regeneration and neuromodulation totreatment of rehabilitation, and to supportfor exploring the neurobiological mechanism of rehabilitation. This review will explore the important role of multi-target synergistic enhanced neural circuit modulation technology in neural circuit reconstruction, and establish more accurate and effective multi-target innovative technologies and clinical pathways, so as to promote the basic and clinical research of neuromodulation.

To investigate the effects of exosomes of mouse astrocytes on the viability of neural stem cells.Cultured and isolated the mouse astrocytes, and collected the cell supernatant for obtain the exosomes by ultracentrifugation. Neural stem cells that primary cultured for 2① Identification of astrocyte exosomes: The successful obtain of exosomes of cell supernatant were confirmed by techniques such as electron microscopy, Western blot, exosome concentration and particle size measurement. ② CCK8 experiment: As the increasement of the concentration of exosomes, cell proliferation of primary neural stem cells gradually increased. Compared with the control group, proliferation of the cells in 40 μg/mL and 60 μg/mL exosome treatment groups was significantly enhanced, but there was no significant difference between the two groups. So, 40 μg/mL was selected as the best intervention concentration. ③ EdU detection: Number of EdU positive labeled cells in the 40 μg/mL exosome group was higher than that in the control group (Mouse astrocyte exosomes can improve the viability of neural stem cells.

This article reports a patient with spinal cord injury who was treated with conventional rehabilitation therapy plus repeated transcranial magnetic stimulation (rTMS) during the postoperative rehabilitation, and to observe the effects of rTMS on dystonia. A 66-year-old male patient fell from the bed 8 months ago. At that time, he felt pain in his neck, dysfunction in limbs movement, and loss of sensation in trunk and limbs. Magnetic resonance imaging (MRI) revealed spinal cord injury. Under general anesthesia, anterior cervical decompression and bone graft fusion (ACDF) and cervical spine internal fixation of C

To study the effect and mechanism of modified constraint-induced movement therapy (mCIMT) on motor function recovery in cerebral ischemia-reperfusion rats.The rats were randomly divided into the control group and the mCIMT group, with 12 rats in each group. The left middle cerebral artery occlusion (MCAO) model was established by the Longa suture method. In the mCIMT group, the rats started continuous training for 14 d on the 7Compared with the control group, the body mass of rats in the mCIMT group was comparable (mCIMT improved the motor function of MCAO rats, and the mechanism might be related to the increase of amino acid neurotransmitters and 5-HIVV and decrease of p-p70s6k expression in the motor cortex.

To provide a new method for investigating the histological characteristics of acupoints by obser-ving the microstructure of the lymphatic vessels in the skin tissue of "Taichong" (LR3) and "Yongquan" (KI1) regions.Six male SD rats were used in the present study. The skin tissue of LR3 and KI1 from the hind foot were taken following transcardial perfusion with 4% paraformaldehyde. The skin tissues were cut into sagittal sections with a freezing microtome and stained by fluorescent immunohistochemistry with lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), calcitonin gene-related peptide (CGRP), and phalloidin for displaying the lymphatic vessels, nerve fibers, and blood vessels, separately. The samples were viewed and recorded using fluorescent microscope and laser scanning confocal microscope.In the skin tissue of LR3 and KI1 regions, the lymphatic vessels, nerve fibers, and blood vessels were labeled with LYVE-1, CGRP and phalloidin, respectively. The lymphatic capillaries were found to start from the enlarged blind end and distribute in the dermis and subcutaneous tissues with various forms, crisscrossing. Abundant blood capillaries at various thickness distributed around the lymphatic capillaries in a parallel or crossed pattern, intermingled with free nerve fibers.The lymphatic capillaries, blood capillaries and nerve fibers extensively distribute in the skin tissues of LR3 and KI1 regions in rats, suggesting an involvement of the immunomodulation in the effects of acupuncture in pathological conditions, despite being not limited to the acupoint regions in the distribution of lymphatic capillaries.

Histological pictures are important parts of medical research articles, and a good histological picture not only visualizes the content of the article, but also makes it easier for readers to understand. With the rapid development of microscopic technology and image processing technology, journals have higher requirements for the quality of histological pictures and more strict requirements for the editing and annotation of pictures. Based on an analysis of the current status of histological pictures in the articles of

Background: Adipose-derived mesenchymal stem cells ( ADMSCs ) have attracted widespread interest as cell-based tissue repair systems. To obtain adequate quantities of ADMSCs for therapeutic applications, extensive in vitro expansion is required. However, under current two-dimensional (2D) approaches , ADMSCs rapidly undergo replicative senescence , and cell growth is impeded and stem cell properties are eliminated by mechanisms that are poorly understood . These issues limit the extensive applications of ADMSCs . In this study, we investigated senescence-related changes in mesenchymal stem cells (MSCs) isolated from human adipose tissue in 2D and three-dimensional (3D) cultures. Methods: We studied cell growth over a given period (21 days) to determine if modes of culture were associated with ADMSCs senescence . ADMSCs were isolated from healthy females by liposuction surgery and then were grew in 2D and 3D cultures. The cell morphology was observed during cell culture. Every other time of culture, senescence-associated β-galactosidase (SA-β-gal) expression, cell viability, proliferation, and differentiation potential of ADMSCs from 2D and 3D cultures were detected. Also, senescence and stemness related genes expression, telomere length, telomerase activity, and energy metabolism of ADMSCs for different culture time were evaluated. Results: With long-term propagation, we observed significant changes in cell morphology, proliferation, differentiation abilities and energy metabolism, which were associated with increases in SA-β-gal activity, and decreases in telomere length and telomerase activity . Notably, when cultured in 3D, these changes were improved. Conclusions: Our results indicate that 3D culture is able to ameliorate senescence-related changes in ADMSCs.

A fibre Bragg grating (FBG)-based load and vibration transducer (FLVT) was developed using a 3D fused deposition modelling (FDM) approach. A newly FLVT was designed by the equal-strength cantilever beam in which the FBG sensors were embedded in the beam during the FDM process. The temperature effect was eliminated by the temperature sensor in the vibration sensing unit. The parameters of the proposed FLVT was examined by the finite element method. The simulated results were matched well with the theoretical analysis results and laboratory calibration results. The proposed transducer has the pressure measurement sensitivity of 0.01274 nm kPa−1 for the earth pressure below 150 kPa. In addition, the proposed transducer could accurately measure low-frequency vibration signals with maximum frequency of 4 Hz and the maximum displacement amplitude of 4 mm with sensitivity of 117.6 pm g−1. The measurement accuracy and stability were carried out. Results shown that the maximum relative errors between the calculation results and the experimental results was 1.3%. The effect of vibration direction was also analysed for the proposed FLVT. The results indicated that the transversal vibration has less influence on the longitudinal vibration. The outcome of this study indicated that the proposed FLVT provide a newly approach for the measurement of earth pressure and soil vibration in one transducer which is quit suit for soil mass.

Activation and reconstruction of the spinal cord circuitry is important for improving motor function following spinal cord injury. We conducted a case series study to investigate motor function improvement in 14 patients with chronic spinal cord injury treated with 4 weeks of unilateral (right only) cortical intermittent theta burst stimulation combined with bilateral magnetic stimulation of L3–L4 nerve roots, five times a week. Bilateral resting motor evoked potential amplitude was increased, central motor conduction time on the side receiving cortical stimulation was significantly decreased, and lower extremity motor score, Berg balance score, spinal cord independence measure-III score, and 10 m-walking speed were all increased after treatment. Right resting motor evoked potential amplitude was positively correlated with lower extremity motor score after 4 weeks of treatment. These findings suggest that cortical intermittent theta burst stimulation combined with precise root stimulation can improve nerve conduction of the corticospinal tract and lower limb motor function recovery in patients with chronic spinal cord injury.

Neural tract tracing is used to study neural pathways and evaluate neuronal regeneration following nerve injuries. However, it is not always clear which tracer should be used to yield optimal results. In this study, we examined the use of Alexa Fluor 488-conjugated cholera toxin subunit B (AF488-CTB). This was injected into the gastrocnemius muscle of rats, and it was found that motor, sensory, and sympathetic neurons were labeled in the spinal ventral horn, dorsal root ganglia, and sympathetic chain, respectively. Similar results were obtained when we injected AF594-CTB into the tibialis anterior muscle. The morphology and number of neurons were evaluated at different time points following the AF488-CTB injection. It was found that labeled motor and sensory neurons could be observed 12 hours post-injection. The intensity was found to increase over time, and the morphology appeared clear and complete 3-7 days post-injection, with clearly distinguishable motor neuron axons and dendrites. However, 14 days after the injection, the quality of the images decreased and the neurons appeared blurred and incomplete. Nissl and immunohistochemical staining showed that the AF488-CTB-labeled neurons retained normal neurochemical and morphological features, and the surrounding microglia were also found to be unaltered. Overall, these results imply that the cholera toxin subunit B, whether unconjugated or conjugated with Alexa Fluor, is effective for retrograde tracing in muscular tissues and that it would also be suitable for evaluating the regeneration or degeneration of injured nerves.

Classic paired associative stimulation can improve synaptic plasticity, as demonstrated by animal experiments and human clinical trials in spinal cord injury patients. Paired associative magnetic stimulation (dual-target peripheral and central magnetic stimulation) has been shown to promote neurologic recovery after stroke. However, it remains unclear whether paired associative magnetic stimulation can promote recovery of lower limb motor dysfunction after spinal cord injury. We hypothesize that the current caused by central and peripheral magnetic stimulation will converge at the synapse, which will promote synapse function and improve the motor function of the relevant muscles. Therefore, this study aimed to examine the effects of paired associative magnetic stimulation on neural circuit activation by measuring changes in motor evoked and somatosensory evoked potentials, motor and sensory function of the lower limbs, functional health and activities of daily living, and depression in patients with spinal cord injury. We will recruit 110 thoracic spinal trauma patients treated in the Department of Spinal Cord Injury, China Rehabilitation Hospital and randomly assign them to experimental and control groups in a 1:1 ratio. The trial group (n = 55) will be treated with paired associative magnetic stimulation and conventional rehabilitation treatment. The control group (n = 55) will be treated with sham stimulation and conventional rehabilitation treatment. Outcomes will be measured at four time points: baseline and 4, 12, and 24 weeks after the start of intervention (active or sham paired associative magnetic stimulation). The primary outcome measure of this trial is change in lower limb American Spinal Injury Association Impairment Scale motor function score from baseline to last follow-up. Secondary outcome measures include changes in lower limb American Spinal Injury Association sensory function score, motor evoked potentials, sensory evoked potentials, modified Ashworth scale score, Maslach Burnout Inventory score, and Hamilton Depression Scale score over time. Motor evoked potential latency reflects corticospinal tract transmission time, while amplitude reflects recruitment ability; both measures can help elucidate the mechanism underlying the effect of paired associative magnetic stimulation on synaptic efficiency. Adverse events will be recorded. Findings from this trial will help to indicate whether paired associative magnetic stimulation (1) promotes recovery of lower limb sensory and motor function, reduces spasticity, and improves quality of life; (2) promotes neurologic recovery by increasing excitability of spinal cord motor neurons and stimulating synaptic plasticity; and (3) improves rehabilitation outcome in patients with spinal cord injury. Recruitment for this trial began in April 2021 and is currently ongoing. It was approved by the Ethics Committee of Yangzhi Affiliated Rehabilitation Hospital of Tongji University, China (approval No. YZ2020-018) on May 18, 2020. The study protocol was registered in the Chinese Clinical Trial Registry (registration number: ChiCTR2100044794) on March 27, 2021 (protocol version 1.0). This trial will be completed in April 2022.

Following a spinal cord injury, there are usually a number of neural pathways that remain intact in the spinal cord. These residual nerve fibers are important, as they could be used to reconstruct the neural circuits that enable motor function. Our group previously designed a novel magnetic stimulation protocol, targeting the motor cortex and the spinal nerve roots, that led to significant improvements in locomotor function in patients with a chronic incomplete spinal cord injury. Here, we investigated how nerve root magnetic stimulation contributes to improved locomotor function using a rat model of spinal cord injury. Rats underwent surgery to clamp the spinal cord at T10; three days later, the rats were treated with repetitive magnetic stimulation (5 Hz, 25 pulses/train, 20 pulse trains) targeting the nerve roots at the L5-L6 vertebrae. The treatment was repeated five times a week over a period of three weeks. We found that the nerve root magnetic stimulation improved the locomotor function and enhanced nerve conduction in the injured spinal cord. In addition, the nerve root magnetic stimulation promoted the recovery of synaptic ultrastructure in the sensorimotor cortex. Overall, the results suggest that nerve root magnetic stimulation may be an effective, noninvasive method for mobilizing the residual spinal cord pathways to promote the recovery of locomotor function.

A quantitative phase field method of multi-component diffusion-controlled phase transformations coupled with the Kim–Kim–Suzuki model was applied to study the effect of initial particle size distribution (PSD) in 3D and space distribution in 2D on dissolution of α particles in Ti–6Al–4V alloy below β transus temperature in real time and length scale. The thermodynamic and mobility data were obtained from Thermo-Calc and DICTRA softwares, respectively. The results show that the volume fractions of α particles decay with time as: $$ f = f_{\text{eq}} + (f_{0} - f_{\text{eq}} )\exp ( - Kt^{n} ) $$ for four cases of PSD. The sequence of dissolution kinetics from fast to slow is: uniform PSD, normal PSD, lognormal PSD and bimodal PSD. The space distribution is found to be a major factor affecting the dissolution kinetics and the microstructures. When the distance of the particles is less than critical value, the dissolution rates reduce with the decrease in distance. The Al and V concentration fields around the particles appear more obvious soft impingement.

To collect literatures on the application of neural tracing technique in experimental acupuncture research, and summarize its application status in acupoints, meridians and Zang-fu organs.We collected papers published from databases of CNKI (1979-2017), Wan-Fang (1990-2017), VIP (1989-2017) and PubMed(1997-2017)by using keywords of "neural tracing technique" "neuroanatomic tracing" "neural tracers" and "acupuncture" "electroacupuncture" "auricular acupuncture" "eye acupuncture" "meridians" "acupuncture points" "acupoint injection", and made a summary about the current state of application of neural tracing technique in the fields of acupoint, meridian and Zang-fu organs (viscera).A total of 94 articles were collected, the most commonly used neural tracers were horseradish peroxidase and cholera toxin subunit B. The experimental animals used were rat, rabbit, cat, monkey, etc., and injection site was acupoint. After the injection of neural tracers, the survival time of animals was range from 1 day to 12 weeks, and the labelled tissues included neurons and nerve fibers of the sensory, motor, and autonomic systems. The outcomes of neural labeling mainly revealed the segmental pattern, neuroanatomical connection (neural pathways/circuits) and chemical features (shown by immunohistochemical staining) of neurons and nerve fibers innervating both the acupoints and visceral organs, suggesting their involvement in the effect of acupuncture and moxibustion treatment.This application of neural tracing technology help us understand the under-lying mechanisms of acupuncture and moxibustion interventions from different perspectives of neural pathways/circuits and related chemical properties, which also lays a greater role for this technology in future experimental acupuncture research.

To investigate the expression of serotonin (5-HT) in local tissue of different acupoint regions at the hind limbs of rats.A total of 6 male SD rats were used in the present study. The local tissues of "Neiting"(ST44), "Zusanli"(ST36) and "Futu"(ST32) at the hindlimbs were dissected after transcardiac perfusion with fixative solution containing 4% paraformaldehyde, phosphate buffer, etc., fixed again in 4% paraformaldehyde and cut into sections (at 20 µm thick). Then, the tissue sections were subjective into immunofluorescence stain for displaying 5-HT positive cells and calcitonin gene-related peptide (CGRP) -positive fibers, vessel-like structure (labelled with phalloidin) and cellular nucleus (labelled with DAPI), separately. After that, the tissue samples were observed and recorded using a laser confocal microscope.The 5-HT labeled cells presented flat or round form in the local tissues of the 3 acupoint regions. These cells were mainly located in the dermis and subcutaneous tissue, and concentrated around the vessel-like structure and CGRP-positive nerve fibers. The number of cutaneous 5-HT positive cells was most in ST44 (156.6±12.1), and fewer in ST36 (83.5±18.2)and ST32(53.3±5.2)in turn.5-HT-positive cell is an important member in local tissue of acupoints (ST44, ST36 and ST32), and varies in number according to their locations, suggesting an involvement of different densities of 5-HT-positive cells in different levels of regulatory effects of acupuncture stimulation of these acupoints.

To explore the effect of moxa-stick moxibustion and joss-stick moxibustion at "Guanyuan" (CV4) on the activity of mast cells in the small intestine tissue in rats.Twelve male SD rats were randomly divided into control, joss-stick moxibustion and moxa-stick moxibustion groups (Compared with the control group, the skin temperature of the CV4 region in both the joss-stick and the moxa-stick groups were significantly increased (Moxa-stick moxibustion gives rise a higher temperature at CV4 to activate mast cells surrounded by CGRP and NPY positive nerve fibers in the small intestine tissue in rats, suggesting an involvement of the sensory and sympathetic nervous system in the activation of intestinal mast cells possibly by way of somatic sympathetic reflex.

To try to re-map an acupoint atlas in vector mode in the experimental rat by using a new-generation drawing tool.A total of 5 SD rats were used in the present study. In reference to the current acupoint locations and anatomical structure of rats as well as those of the human body, an acupoint atlas having a stereoscopic mode was re-mapped by using Adobe photoshop/illustrator CS6 imaging processing and drawing system.According to the skeletal structure and body surface landmarks, an acupoint atlas consisting of Shuigou (GV26), Baihui (GV20), Tianmen (i.e. Cuanzhu, BL2), Erjian (MA-H6), Dazhui (GV14), Feishu (BL13), Xinshu (BL15), Geshu (BL17), Jizhong (GV6), Pishu (BL20), Shenshu (BL23), Houhui (SI3), Huantiao (GB30), Houhai, Yanglingquan (GB34), Housanli (ST36), Zhaohai (KI6), Sanyinjiao (SP6), Genduan point, Shenmai (BL62), Taichong (LR3), Zhijian (foot), Yongquan (KI1), Guanyuan (CV4), Xiqian, Weijian, Shenque (CV8), Zhongwan (CV12), Qiansanli (LI10), Waiguan (SJ5), Neiguan (PC6), Quchi (CV6), Zhoujie (EX-UE1), Hegu (LI4), Zhijian (hand), Houxi (SI3), Shenmen (HT7), Taiyuan (LU9), Shaohai (HT3), Chize (LU5), Danzhong (CV17), and Chengjiang (CV24) which have been positioned in plane graph mode by Professor HUA Xing-bang and colleagues in 1991. The newly re-mapped acupoint atlas displayed more clearly in the anatomic locations relevant to the traditional one.A new acupoint atlas with 3 dimension image mode is accomplished in the rat, being more convenient and applicable for researchers to edit the relevant graph materials in paper writing, and playing a possibly useful guidance for the standardization of acupoint selection in experimental acupuncture researches.

Biotinylated dextran amine (BDA) has been used for neural pathway tracing in the central nervous system for many decades, in which high molecular weight BDA appeared to be transported predominantly in the anterograde direction and less in the retrograde direction. In the current study, we reexamined the properties of neural labeling with high molecular weight BDA through a reciprocal neural pathway between thalamus and somatosensory cortex. After injection of BDA into the ventral posteromedial nucleus of thalamus (VPM) in the rat, the BDA labeling was sequentially examined on somatosensory cortex at 3, 5, 7, 10, and 14 survival days. Both of anterogradely labeled axonal terminals and retrogradely labeled neuronal cell bodies were observed simultaneously on the somatosensory cortex. With the increasing of survival times after injection, morphological changes occurred on the labeled axonal arbors and neuronal dendrites, in which the high quality of BDA labeling appeared on the tenth survival day. These results indicate that high molecular weight BDA is not only a sensitive anterograde tracer but also an excellent retrograde marker to be used for tracing through thalamocortical and corticothalamic pathways. And the detailed structure of neural labeling with BDA similar to Golgi-like resolution can be obtained at optimal survival times of animals after the injection of high molecular weight BDA.

The incidence of severe traumatic brain injury (TBI) is gradually increasing, and its integrated treatment and rehabilitation in early-stage is increasingly valued by working together with doctors in surgical intensive care unit (SICU), surgeons and rehabilitation doctors. In this report, we presented a seven-year-old child was hit on the right side of the head by a falling glass from the building, causing severe TBI, starting from emergency room (ER), SICU, neurosurgery, spine surgery, whole–process and intensive rehabilitation. The child functionally regained some of his motor functions and most of his cognitive functions step by step from coma to cognition and was expected to return to his schooling finally. The authors of this report discuss the necessity and effectiveness of multidisciplinary integration and early intervention in rehabilitation. The case exhibited that multiple rehabilitation procedures would be conducted in whole process of patients with trauma. Furthermore, rehabilitation physician and therapist play critical roles in evaluation and handling of treatment strategy for precise movement and cognitive functional recovery. Particularly, the cognitive function assessment and treatment has significantly performed in making decision on the opportunity of operation of hydrocephalus. All above, the case offered the model of the integrated rehabilitation in severe TBI.

Laser scanning confocal microscope (LSCM) has been used in the field of morphological research of acupuncture and moxibustion for more than 20 years. It is one of the important tools for revealing the structure of acupoints and the effect of acupuncture from histological perspective. With the help of technical advantages of LSCM, the quality of morphological research of acupuncture and moxibustion has been greatly improved, helping us gain a deeper understanding about the structure of acupoints and meridians as well as histochemical changes induced by acupuncture/moxibustion intervention. In order to promote the application of LSCM in acupuncture and moxibustion, we simply reviewed some recent studies in this field and combined them with our experience, trying to provide some technical suggestions. We expect that the technique of LSCM could be integrated into more experiments in acupuncture medicine to provide more powerful morphological evidence for exploring the underlying mechanisms of acupuncture and moxibution therapies.

To explore the neuroanatomical basis of acupoint-visceral correlation by studying the distribution of the neurons associated with acupoint "Shenshu" (BL 23) area and adrenal gland in rats.AF 488-CTB and AF 594-CTB were injected into the left side of BL 23 area and adrenal gland in the same rat respectively. Three days after injection, the dorsal root ganglions (DRG), sympathetic chain, and spinal cord were dissected out from the perfused rats. The neuronal labeling with AF 488/594-CTB was directly observed on the sections under a fluorescent microscope or a laser scanning confocal microscope.All neural labeling was observed in the injection side. The sensory neurons associated with both acupoint BL 23 and adrenal gland distributed from thoracic (T) 10 to lumbar (L) 2 DRG with high concentration in T 12-T 13 and T 11-T 12, respectively, in which some of them were simultaneously labeled with both AF 488/594-CTB and located in T 12-L 1 DRG. For the sympathetic innervation, the postganglionic neurons correlated with BL 23 and adrenal gland were labeled with AF 488/594-CTB separately in the sympathetic chain at the lumbar segments, while the labeled preganglionic neurons were only observed at the lateral horn of T 11-T 13 spinal segments in the cases of adrenal gland. In addition, the labeled motor neurons were mainly detected in the spinal ventral horn at cervical (C) 7-C 8 and T 11-L 1 segments.These results indicate that there are segmental correlation between BL 23 and adrenal gland on the sensory and sympathetic innervations, and this correlation might be a neural pathway for modulating the function role of adrenal gland through BL 23 needling.

The anatomical structure of acupuncture points is the carrier for needling stimulation induced neurobiological effects. Although researchers have never stopped investigating its structure and experienced different stages from macroscopic to microcosmic morphological material basis, our understanding still largely lags behind the requirements of the times. In the present paper, we review development of researches on acupuncture points in recent decades from 1) macroscopic anatomical structure including skin, bones, muscles, nerves, blood vessels, etc.; 2) macro-microcosmic structure as the thickness of skin and muscles, distribution of blood vessels and nerve fibers, and innervation; 3) microstructure including cell categories (mast cells, fibroblasts, macrophages, etc.), peptidergic nerve fibers, various receptors, chemical substances, and their relationship with the brain and immune-endocrine system (cutaneous-neuro-immune-endocrine-mental biological communication), etc. These research outcomes will not only help us demonstrate the acupoint's structure better at the level of the body surface and regional anatomy, but also, at the microstructural level, reveal different kinds of cells and their chemical compositions in its structure as well as their relationship with the regulatory system of the whole body. From the perspective of the cutaneous and systemic neuro-immuno-endocrine networks, it undoubtedly prompts us to profoundly explore the underlying biological mechanisms of acupuncture. To observe the regional mutual connection and interaction of different kinds of cells and their chemical mediators and molecules in the acupuncture point will improve our understanding about the nature of acupuncture points from the static to the dynamic point of view. This might be a new approach for us to further reveal the functional roles of acupuncture points.

High molecular weight biotinylated dextran amine (BDA) has been used as a highly sensitive neuroanatomical tracer for many decades. Since the quality of its labeling was affected by various factors, here, we provide a refined protocol for the application of high molecular weight BDA for studying optimal neural labeling in the central nervous system. After stereotactic injection of BDA into the ventral posteromedial nucleus (VPM) of the thalamus in the rat through a delicate glass pipette, BDA was stained with fluorescent streptavidin-Alexa (AF) 594 and counterstained with fluorescent Nissl stain AF500/525. On the background of green Nissl staining, the red BDA labeling, including neuronal cell bodies and axonal terminals, was more distinctly demonstrated in the somatosensory cortex. Furthermore, double fluorescent staining for BDA and the calcium-binding protein parvalbumin (PV) was carried out to observe the correlation of BDA labeling and PV-positive interneurons in the cortical target, providing the opportunity to study the local neural circuits and their chemical characteristics. Thus, this refined method is not only suitable for visualizing high quality neural labeling with the high molecular weight BDA through reciprocal neural pathways between the thalamus and cerebral cortex, but also will permit the simultaneous demonstration of other neural markers with fluorescent histochemistry or immunochemistry.

Objectives To compare the neural pathways associated with the tissues located at different traditional acupuncture points in the rat forehead and face using the cholera toxin B subunit (CTB) neural tracing technique. Methods After injection of CTB into the tissues at GB14, ST2 and ST6 in the rat, the neural labelling associated with each acupuncture point was revealed by fluorescent immunohistochemistry of the nervous system, including the trigeminal ganglion (TRG), cervical dorsal root ganglia (DRG), spinal cord and brain. Results The CTB labelling included sensory neurons and their transganglionic axonal terminals, as well as motor neurons. The labelled sensory neurons associated with GB14, ST2 and ST6 were distributed in both the TRG and cervical DRG, and their centrally projected axons terminated in an orderly fashion at their corresponding targets in the spinal trigeminal nucleus and cervical spinal dorsal horn. In addition, labelled motor neurons were observed in the facial motor nucleus, trigeminal motor nucleus and cervical spinal ventral horn, in which facial motor neurons projected to the tissues located at all three acupuncture points. Trigeminal motor neurons innervated both ST2 and ST6, while spinal motor neurons only correlated with ST6. Conclusions These results indicate that the tissues located at each of these three traditional acupuncture points in the rat forehead and face has its own sensory and motor connection with the nervous system in a region-specific pattern through distinct neural pathways. Understanding the neuroanatomical characteristics of acupuncture points from the peripheral nervous system to the central nervous system should help inform acupuncture point selection according to the demands of the clinical situation.

Previous studies have shown that the effects of manual acupuncture (MA) are contributed by collagen fibers and mast cells in local acupoints, at which acupuncture stimulation causes various afferent fiber groups to be excited. However what happens in local nerve fibers and mast cells after MA remains unclear. The aim of this study was to examine the response of cutaneous nerve fibers and mast cells to MA stimulation in acupoint Hegu (LI4). The contralateral LI4 of the same rat was used as a non-stimulated control. Immnohistochemistry analysis were carried out to observe the expression of histamine (HA), serotonin (5-HT) and nociceptive neuropeptides, calcitonin gene-related peptide (CGRP) and substance P (SP), in the LI4 area. Mast cells were labeled with anti-mast cell tryptase antibody and simultaneously with HA or 5-HT primary antibodies to observe their co-expression. Our results showed that SP and CGRP were expressed more highly on the cutaneous nerve fibers of LI4 after MA stimulation than that of the control. Mast cells aggregated in close proximity to the blood vessels in intra-epidermis and dermis and some of them with degranulation in the lower dermis and subcutaneous tissue of LI4. Both mast cells and their granules appeared with HA (+) and 5-HT (+) expression at stimulated L14 sites, while a few intact mast cells with a little expression of 5-HT and HA were distributed in areas of non-stimulated L14. The results indicated that local cutaneous nerve terminals and mast cells responded to MA with higher expression of SP and CGRP in nerve fibers, as well as with aggregation and degranulation of mast cells with HA and 5-HT granules at acupoint LI4. These neuroactive substances may convey signals to certain pathways that contribute to the effects of acupuncture.

According to the objective requirements of teaching transformation and the history of school curriculum construction, we further clarify the main factors affecting the quality of curriculum teaching, center on the construction and reform of the curriculum, clarify the goals and tasks of deepening the construction and reform, and put forward the specific measures to promote the reform Measures. After more than two years of deepening the practice of curriculum construction and reform, the curriculum system is being reconstructed, the curriculum projects are progressing well, and the curriculum team is gradually formed. It is foreseeable that continuing the implementation of the strategic initiatives of deepening curriculum construction and reform in schools will effectively support the teaching transformation in schools so as to make outstanding contributions to the school's strategic goal of "transformation and upgrading".