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    Neurobiology

    Characterization of group Ⅰ metabotropic glutamate receptors in rat superior cervical ganglion and their changes following chronic intermittent hypoxia

    WEI Xi-xi LI Chao-hong ZHAO Chen-lu TANG Jia-ping LIU Yu-zhen
    2024, 55 (1):  3-9.  doi: 10.16098/j.issn.0529-1356.2024.01.001
    Abstract ( )   PDF (12732KB) ( )  

    Objective  To observe the expression and localization of group Ⅰ metabotropic glutamate receptors (mGluR1/5) in rat superior cervical ganglion (SCG) and the effect of chronic intermittent hypoxia (CIH) on mGluR1/5 protein level.   Methods  Twelve male SD rats were randomly divided into control group(Ctrl)and CIH group(CIH), 6 rats in each group. After 6 weeks of modeling, the effect of CIH on mGluR1/5 protein level was detected by Western blotting, the expression and distribution of mGluR1/5 in SCG were detected by immunohistochemistry and double-immunofluorescent staining.     Results  mGluR1/5 was expressed in rat SCG. mGluR1 was distributed in neurons and small intensely fluorescent (SIF) cells, but not in satellite glial cells (SGCs), nerve fibers and blood vessels, whereas mGluR5 was mainly distributed in nerve fibers and a little in neurons, but not in SGCs, SIF cells and blood vessels. CIH increased the protein levels of mGluR1/5 (P<0.01) in rat SCG.   Conclusion  Both mGluR1 and mGluR5 are expressed in the rat SCG, but their distribution are different, and the increased protein levels of both may be involved in CIH-induced hypertension.

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    Expression and distribution of brain-derived neurotrophic factor in different cerebrum regions of yak and cattle

    ZHENG Li-ping LIU Xia DU Xiao-hua WU Ya-juan LIU Shan-shan
    2024, 55 (1):  10-16.  doi: 10.16098/j.issn.0529-1356.2024.01.002
    Abstract ( )   PDF (10237KB) ( )  

    Objective  To clarify the expression and distribution of brain-derived neurotrophic factor (BDNF) in the cerebrum of plateau yaks and cattle, and to explore the relationship between BDNF function and the adaptability of altitude hypoxia.    Methods  Five yaks and five cattles were selected.The content and distribution of BDNF in frontal lobe, temporal lobe, parietal lobe, occipital lobe, cerebrum white matter and hippocampus of yak and cattle were analyzed by Real-time PCR, Western blotting and Immunohistochemistry.   Results  Real-time PCR result  showed that BDNF mRNA expression in the cerebrum of yaks and cattles was highest in temporal cortex, followed by hippocampus, parietal cortex, occipital cortex and frontal cortex, and lowest in white matter.  Western blotting results showed that the content of BDNF protein in the cerebrum of yaks was the highest in temporal cortex,followed by hippocampus. The content of BDNF protein in other tissues was parietal cortex, frontal cortex and cerebrum white matter, and the content of BDNF protein was the lowest in occipital cortex. The content of BDNF protein intlecerebrum of cattles was the highest in the temporal cortex, followed by the hippocampus. The content of BDNF protein in other tissues was parietal cortex, occipital cortex and frontal cortex in descending order, and the protein content in cerebrum white matter was the lowest.  Immunohistochemical results showed that the positive expression of BDNF protein in the cerebrum of yaks and cattles was basically similar, mainly distributed in the granulosa cells and glial cells in the frontal cortex, temporal cortex, parietal cortex and occipital cortex, glial cells in cerebrum white matter, pyramidal cell layer and polyform cell layer in the hippocampus. There was the small amount of distribution in Martinotti cells and the molecular layer of hippocampus in the cerebral cortex.   Conclusion  BDNF mRNA and protein are distributed and expressed in different brain regions of yaks and cattles, but the expression level different, which is speculated to be closely related to the specific functions of different cerebrum regions. The expression level of the cerebrum of yak is higher than that of cattle except occipital cortex, suggesting that it is related to the altitude hypoxic environment. BDNF may play an important role in enhancing hypoxic tolerance and protecting internal environmental homeostasis in the process of animal adaptation to hypoxic environment.

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     Analysis of cerebral gray matter structure in multiple sclerosis and neuromyelitis optica

    LIU Xiao-li XU Lin WU Ai-xue WEN Cai-yun LI Ru-hua WU An-ting CHEN Dai-qian CHEN Cheng-chun
    2024, 55 (1):  17-24.  doi: 10.16098/j.issn.0529-1356.2024.01.003
    Abstract ( )   PDF (9030KB) ( )  

    Objective  The volume and cortical thickness of gray matter in patients with multiple sclerosis (MS) and neuromyelitis optica (NMO) were compared and analyzed by voxel based morphometry (VBM) and surface-based morphometry (SBM), and the differences in the structural changes of gray matter in the two diseases were discussed.   Methods  A total of 21 MS patients, 16 NMO patients and 19 healthy controls were scanned by routine MRI sequence. The data were processed and analyzed by VBM and SBM method  based on the statistical parameter tool SPM12 of Matlab2014a platform and the small tool CAT12 under SPM12.   Results  Compared with the normal control group (NC), after Gaussian random field (GRF) correction, the gray matter volume in MS group was significantly reduced in left superior occipital, left cuneus, left calcarine, left precuneus, left postcentral, left central paracentral lobule, right cuneus, left middle frontal, left superior frontal and left superior medial frontal (P<0.05). After family wise error (FWE) correction, the thickness of left paracentral, left superiorfrontal and left precuneus cortex in MS group was significantly reduced (P<0.05). Compared with the NC group, after GRF correction, the gray matter volume in the left postcentral, left precentral, left inferior parietal, right precentral and right middle frontal in NMO group was significantly increased (P<0.05). In NMO group, the volume of gray matter in left middle occipital, left superior occipital, left inferior temporal, right middle occipital, left superior frontal orbital, right middle cingulum, left anterior cingulum, right angular and left precuneus were significantly decreased (P<0.05). Brain regions showed no significant differences in cortical thickness between NMO groups after FWE correction. Compared with the NMO group, after GRF correction, the gray matter volume in the right fusiform and right middle frontal in MS group was increased significantly(P<0.05). In MS group, the gray matter volume of left thalamus, left pallidum, left precentral, left middle frontal, left middle temporal, right pallidum, left inferior parietal and right superior parietal were significantly decreased (P<0.05). After FWE correction, the thickness of left inferiorparietal, left superiorparietal, left supramarginal, left paracentral, left superiorfrontal and left precuneus cortex in MS group decreased significantly (P<0.05).   Conclusion  The atrophy of brain gray matter structure in MS patients mainly involves the left parietal region, while NMO patients are not sensitive to the change of brain gray matter structure. The significant difference in brain gray matter volume between MS patients and NMO patients is mainly located in the deep cerebral nucleus mass.

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    Neurobiology

    Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice

    YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei
    2024, 55 (1):  25-31.  doi: 10.16098/j.issn.0529-1356.2024.01.004
    Abstract ( )   PDF (4060KB) ( )  

     Objective To analyse the analgesic effect and possible mechanism of panax notoginseng saponin (PNS) on mouse models of chronic inflammatory pain caused by complete Freund’s adjuvant (CFA).   Methods A total of 48 male C57BL/6J mice were divided randomly into four groups: normal saline control group (Ctrl), CFA group (CFA), CFA + PNS group (CFA+PNS), CFA + dexamethasone (DEX) group (CFA+DEX). Von Frey filaments were used to detect mechanical pain in mice. Immunohistochemistry was used to detect the number and morphological changes of glial fibrillary acidic protein (GFAP) positive astrocytes. Western blotting was used to detect the expressions of GFAP, nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in mice’s spinal cord segments in each group.   Results Compared with the Ctrl group, mice in the CFA group showed a significant decrease in mechanical pain thresholds at day 1, day 3, day 5, day 7, and day 14. Additionally, there was a significant decrease in NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of the mice. PNS intervention could relieve mechanical pain and down-regulate the expressions of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of mice, with no significant difference compared with the CFA+DEX group. CFA group mice had significantly more GFAP positive cells in their posterior horns than Ctrl group mice, as measured by immunohistochemistry; PNS intervention decreased the number of GFAP positive cells in the posterior horn of the spinal cord in model mice;DEX had no effect on the number of GFAP positive cells in the dorsal horn of spinal cord. According to Western blotting results, GFAP expression in the spinal cord of the CFA group was significantly more than that of the Ctrl group; PNS intervention significantly reduced GFAP expression in the spinal cord of CFA group mice;DEX had no effect on the expression of GFAP in the posterior horn of spinal cord.   Conclusion PNS has a good alleviating effect on inflammatory pain, and its mechanism may be related to inhibition of astrocyte activation and NLRP3 inflammasome activation. 

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    Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease

    MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning
    2024, 55 (1):  32-42.  doi: 10.16098/j.issn.0529-1356.2024.01.005
    Abstract ( )   PDF (3561KB) ( )  

    Objective  To investigate the relieving effects of knockdown of long non-coding RNA(lncRNAtaurine up-regulated gene 1 (TUG1) on inhibiting nucleotide binding oligomerization domain like receptor protein 1(NLRP1) inflammasome and the progression of Alzheimers disease.    Methods  Wild-type (WT group,10 mice) or amyloid precursor protein (APP)/presenilin-1 (PS1) transgenic mice (30 mice) with a genetic background of C57/BL6 aged 9-10 weeks were used in this study. APP/PS1 transgenic mice were randomly divided into model group, model+lncRNA TUG1 short hairpin RNA (shRNA) group and model+shRNA non target(NT)group (n=10). Blood samples, cerebral cortex tissues, primary microglial cells and primary astrocytes were collected from mice 12 weeks of age on day 1 (3-month-old) and 32 weeks of age on day 1 (8-month-old), with 5 mice per group at each time point. Real-time PCR analysis was used to detect the expression levels of lncRNA TUG1 and macrophage migration inhibitory factor (MIF) mRNA in cerebral cortex tissues and primary microglial cells, and C1r and C1s mRNA levels in primary astrocytes of 3-month-old and 8-month-old mice in the above 4 groups, respectively. ELISA was used to determine the MIF in plasma samples of the above 4 groups of mice. Primary microglia and astrocytes from the cerebral cortex of 3-month-old and 8-month-old mice were co-cultured. CCK-8 method  was used to determine the proliferation ability of the above cells. Western blotting was used to determine the expression levels of MIF, pro interleukin-1β(pro-IL-1β), apoptosis associated speck-like protein containing a caspase recrult domain(ASC), Caspase-1 (p20), Caspase-1 (full), NLRP1 and NLRP3 in cerebral cortex tissues of 3-month-old and 8-month-old mice. Immunofluorescent staining was used to determine amyloid beta(Aβ)in cerebral cortex of 8-month-old mice.    Results  At the age of 3-month-old and 8-month-old, compared with the WT group, the relative expression level of lncRNA TUG1 and MIF in cerebral cortex tissues and primary microglia of model group mice was significantly up-regulated, with primary microglial cells and astrocytes proliferation ability enhanced (P<0.05). Compared with the model group, the relative expression level of lncRNA TUG1 and MIF cerebral cortex tissues and primary microglia of model+lncRNA TUG1 shRNA group were significantly down-regulated, with primary microglial cells and astrocytes proliferation ability decreased (P<0.05). Compared with the WT group, MIF factor in the peripheral plasma of model group increased significantly, with pro-IL-1β,ASC,Caspase-1 (p20),Caspase-1 (full), NLRP1 and NLRP3 expression level up-regulated in the model group mice cerebral cortex tissues, with increased Aβ immunofluorescent indensity (P<0.05). Compared with the model group, MIF factor in the peripheral plasma, and pro-IL-1β,ASC,Caspase-1 (p20),Caspase-1 (full) and NLRP1 expression in the model+lncRNA TUG1 shRNA group mice cerebral cortex tissues were down-regulated, and Aβ immunofluorescent indensity decreased (P<0.05), while NLRP3 expression level were not changed (P>0.05). There was no significant difference between the model group and the model+shRNA NT group mice of all the above factors (P>0.05).   Conclusion  In APP/PS1 transgenic mice, up-regulation of lncRNA TUG1 and MIF are positively associated with the activation of NLRP1 inflammasome in mice cerebral cortex tissues and primary microglia. Knock-down of lncRNA TUG1 can ameliorate the progression of Alzheimer’s disease.

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    Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury
    ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan
    2024, 55 (1):  43-48.  doi: 10.16098/j.issn.0529-1356.2024.01.006
    Abstract ( )   PDF (4802KB) ( )  

    Objective  To explore the dynamic changes and mechanisms of neurological and cognitive functions in mice with traumatic brain injury (TBI).    Methods  Totally 60 12-month-old Balb/c mice were divided into control group (10 in group) and TBI group (50 in group).TBT model mice were divided into 5 subgroups according to the time of model construction, including model 1 day, model 1 day, model 3 day, model 7 day, model 14 days and model 28 days group with 10 in each group. At the 29th day of the experiment, neurological scores and step down tests were carried out. After the test, the mice were sacrificed for brains which were detected by immunohistochemistry staining, inflammatory cytokine tests and Western blotting.    Results  Compared with the control group, the neurological scores of mice in TBI group increased, and then decreased after the 7th day when the scores reached the peak. However, the latency of step down errors was lower than control group, and the number of step down errors was higher than control group which had no changes. Compared with the control group, the expression of lonized calcium-binding adapter molecule 1(IBA1), chemokine C-X3-C-motif ligand1(CX3CL1), C-X3-C chemokine receptor 1(CX3CR1), NOD-like receptor thermal protein domain associated protein 3(NLRP3), and phosphorylation nuclear factor(p-NF)-κB in TBI group increased and reached to the peak at the 7th day, and then started to decrease. At the same time, the levels of inflammatory cytokines interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) first increased to the peak, and then began to decrease. However, compared with the control group, the expression of amyloid β(Aβ) protein and p-Tau protein in the model group continued to increase at all time.   Conclusion  The TBI model caused continuous activation of microglia along with inflammatory response, which first increased and then decreased, resultsing in neurological scores changes. In addition, the inflammatory response may act as a promoter of Aβ protein deposition and Tau protein phosphorylation, leading to cognitive impairment in mice.

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    Effects of tricholoma matsutake polysaccharides on 1-methy-4-pehnyl-pyridine ion-induced PC12 cell damage

    LÜ Hai-yan SHEN Xi-ya ZHAO Fu-sheng ZHU Mei
    2024, 55 (1):  49-54.  doi: 10.16098/j.issn.0529-1356.2024.01.007
    Abstract ( )   PDF (8211KB) ( )  

    Objective  To investigate the protective mechanism of tricholoma matsutake polysaccharides(TMPagainst 1-methy-4-pehnyl-pyridine ion (MPP+)-induced PC12 cell damage.     Methods  An Parkinson’s disease (PD) PC12 cell model in vitro was constructed using MPP+, and PC12 cell were randomly divided into 5 groups: control group(Ctrl), model group(MDL), TMP high concentration (250 μg/L) group (TMP-H), TMP medium concentration (50 μg/L) group (TMP-M), and TMP low concentration (10 μg/L)group (TMP-L), 3 complex wells in each group. Matsutake polysaccharide treated PC12 cells for 24 hours, 250 μmol/L MPP+ induced PC12 cells were prepared for 24 hours to prepare PD cell models. The cell viability of each group of PC12 cell was detected by CCK-8 method, the lactate dehydrogenase (LDH) release of each group of PC12 cell was detected by LDH method, mitochondrial activity was detected by fluorescence microscopy, the average length of mitochondrial network branches and the number of network branches were calculated by Image J 1.53 r software, the mitochondrial membrane potential was detected by JC-1 method, and the protein Bcl-2/Bax ratio was detected by Western blotting method.    Results  Compared with the Ctrl group, the survival rate of PC12 cell in the MDL group decreased, the LDH release level increased, and the mitochondrial membrane potential (MMP,  ΔΨm) and Bcl-2/Bax ratio decreased (all P<0.05). Compared with the MDL group, the cell survival rate of matsutake polysaccharide in the TMP-H group and the TMP-M group increased, the LDH release level decreased, the ΔΨm and Bcl-2/Bax ratio increased (P<0.05), mitochondrial activity increased, the average length of mitochondrial network branches and the number of network branches increased, and the antagonistic membrane potential decreased (P<0.05), among which the matsutake polysaccharide group was more obvious in the reduction of antagonistic membrane potential.    Conclusion  TMP have a protective effect against MPP+-induced PC12 cell damage, possibly by improving cell viability, reducing LDH release, antagonizing the reduction of mitochondrial membrane potential, and repairing mitochondrial network morphology, thereby reducing apoptosis.

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    Cell and Molecules Biology
    Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis
    HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui
    2024, 55 (1):  55-61.  doi: 10.16098/j.issn.0529-1356.2024.01.008
    Abstract ( )   PDF (11420KB) ( )  

    Objective  To investigate the relationship between nuclear factor(NF)-κB signaling pathway and gender differences in alcoholic liver fibrosis.    Methods  C57BL/6 N mice at 7-8 weeks of age were randomly divided into: male normal group, male model group, female normal group and female model group of 20 mice each. The normal group was fed with control liquid diet for 8 weeks, and the model group was fed with alcoholic liquid diet for 8 weeks combined with 31.5% ethanol gavage (5g/kg twice a week) to establish an alcoholic liver fibrosis model. The mice were executed at the end of 8 weekends, and the alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity, estradiol (E2) and testosterone (T) levels were detected in each group, and the fibrosis was detected in each group by Sirius red staining, and liver histopathological changes were observed by HE staining. The postive area rate of NF-κB-P65 and α-SMA were detected by immunohistochemistry and the expression levels of collagen , tumor necrosis factor-α(TNF-α),phosphorylated nuclear factor κB-P65(p-NF-κB-P65), inhibitor of NF-κB(IκBα), and phosphorylated IκBα(p-IκBα) were detected by wes Western blotting.   Results  The male model group showed increased levels of ALT, AST enzyme activity and T, decreased levels of E2, increased collagen fibrils, increased HE staining necrosis score, increased positive area rate of NF-κB-P65 and α-smooth muscle actin(α-SMA), increased expression of collagen Ⅰ, TNF-α, p-NF-κB-P65, p-IκBα and decreased expression of IκBα.   Conclusion  Alcohol is more likely to cause liver fibrosis in male mice, which may be related to enhanced IκBα phosphorylation releasing NF-κB, contributing to increased phosphorylated NF-κB-P65 and further activation of NF-κB signaling pathway.

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     Association between index finger and ring finger length ratios and polymorphism of homeobox A11 gene locus among Ningxia college students

    YANG Meng-yi NIU Shi-bo ZHANG Jing DANG Jie MA Zhan-bing LU Hong HUO Zheng-hao
    2024, 55 (1):  62-66.  doi: 10.16098/j.issn.0529-1356.2024.01.009
    Abstract ( )   PDF (786KB) ( )  

    Objective  To investigate the association between the index finger and ring finger length ratio (2D∶4D) and of four loci (rs6461992, rs6968828, rs7801581, rs17427875) polymorphism of homeobox (HOX) A11 gene among Ningxia college students.   Methods  Digit camera was used to collect frontal hand photos of 667 Han college students (348 males and 319 females) from Ningxia province; Image analysis software was used to mark the anatomical points and measure finger lengths of the index and ring fingers of both hands; multiplex PCR was used to detect each locus polymorphisms of HOXA11 gene; statistical software was used to compare and analyze the differences and associations of 2D∶4D and gene polymorphisms between different genders.   Results  Among Ningxia Han college students, both left hand and right hand 2D∶4D were significantly higher in females than those of in males (all P<0.05), and there were no significant sex differences in right-left hand 2D∶4D; the genotypes and allele frequencies of rs7801581 locus of HOXA11 gene differed significantly between genders (all P<0.05), and none of the other locus polymorphisms showed any significant sex differences; only female left hand 2D∶4D was significantly associated with rs6461992 locus genotype in the relationship between 2D∶4D and HOXA11 polymorphisms (P<0.05).   Conclusion  There were significant sex differences in 2D∶4D among Han college students in Ningxia, and the rs6461992 locus polymorphism of HOXA11 gene may be associated with the formation of 2D∶4D in females.

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    Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells
    XIE Hui-lan FANG Fang LIN Yi
    2024, 55 (1):  67-72.  doi: 10.16098/j.issn.0529-1356.2024.01.010
    Abstract ( )   PDF (4367KB) ( )  
     Objective  To explore the effect and mechanism of Chir99021 on osteogenic differentiation of rat dental pulp stem cells.    Methods  Primary rat dental pulp stem cells were isolated from rat dental pulp and verified by fluorescence immunoassay. Different concentrations of Chir99021 were set, and the cell proliferation was detected by CCK-8 to select the optimal concentration. Osteogenic differentiation was detected by alizarin red staining. The expression of osteogenic differentiation related genes and proteins recombinant wingless type MMTV integration site famity member 1(Wnt1), Wnt3a and Wnt3a β-expression of catenin, axis inhibition protein 2(Axin 2), dentin sialophosphoprotein(OCN) and dentin matrix acidic phosphoprotein 1(DMP1) was detected by Real-time PCR and Western blotting.    Results  The positive expression of dentin sialophosphoprotein(DSPP) and vimentin indicated that rat dental pulp stem cells were successfully isolated. After osteogenic induction of rat dental pulp stem cells, calcium deposits significantly increased with the addition of glycogen synthase kinase-3β(GSK-3β)  inhibitor Chir99021, calcium deposits were significanted reduced. After osteogenic differentiation of rat dental pulp stem cells, the expression of Wnt1, Wnt3a, β-catenin, Axin2, OCN and DMP1 increased, while the expression of Wnt1, Axin2, OCN and DMP1 decreased with the addition of Chir99021.    Conclusion  Chir99021 can inhibit the osteogenic differentiation of rat dental pulp stem cells after 7 days of induction.
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    Anatomy
     3D hippocampal segmentation based on spatial and frequency domain features adaptive fusion and inter-class boundary region enhancement
    BAI He TENG Ye FENG Lei MENG Hai-wei1 TANG Yu-chun LIU Shu-wei
    2024, 55 (1):  73-81.  doi: 10.16098/j.issn.0529-1356.2024.01.011
    Abstract ( )   PDF (1877KB) ( )  
     Objective  Hippocampal atrophy is a clinically important marker for the diagnosis of many psychiatric disorders such as Alzheimer’s disease, so accurate segmentation of the hippocampus is an important scientific issue. With the development of deep learning, a large number of advanced automatic segmentation method  have been proposed. However, 3D hippocampal segmentation is still challenging due to the effects of various noises in MRI and unclear boundaries between various classes of the hippocampus. Therefore, the aim of this paper is to propose new method  to segment the hippocampal head, body, and tail more accurately.   Methods  To overcome these challenges, this paper proposed two strategies. One was the spatial and frequency domain features adaptive fusion strategy, which reduced the influence of noise on feature extraction by automatically selecting the appropriate frequency combination through fast Fourier transform and convolution. The other was an inter-class boundary region enhancement strategy, which allowed the network to focus on learning the boundary regions by weighting the loss function of the boundary regions between each class to achieve the goal of pinpointing the boundaries and regulating the size of the hippocampal head, body and tail.
       Results  Experiments performed on a 50-case teenager brain MRI dataset show that our method  achieves state-of-the-art hippocampal segmentation. Hippocampal head, body and tail had been improved compared to the existing method. Ablation experiments demonstrated the effectiveness of our two proposed strategies, and we also validated that the network had a strong generalization ability on a 260-case Task04_Hippocampus dataset. It was shown that the method  proposed in this paper could be used in more hippocampal segmentation scenarios.    Conclusion  The method  proposed in this paper can help clinicians to observe hippocampal atrophy more clearly and accomplish more accurate diagnosis and follow-up of the condition.
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    Distal tibiofibular syndesmosis fibular notch typing and its clinical significance based on CT
    YIN Shi-qin YANG Si-yi WANG Rui-han YOU Gui-xuan YANG Ying-qiu ZHANG Lei
    2024, 55 (1):  82-87.  doi: 10.16098/j.issn.0529-1356.2024.01.012
    Abstract ( )   PDF (3769KB) ( )  
     Objective To investigate the morphological typing and clinical significance of the distal tibiofibular syndesmosis fibular notch based on CT images.    Methods  According to the inclusion and exclusion ceiteria, the imaging data of patients undergoing ankle joint CT examination were analyzed, and the inferior tibiofibular joint fibula notch was classified according to the morphological characteristics. The measurements included 8 distances. There were 123 males and 102 females, all of whom were Han nationality, aged 18-60 years old.   Results  Retrospectively analyzed the result  of 225 patients from December 2013 to December 2022. The distal tibiofibular syndesmosis fibular notch was divided into four types according to morphological characteristics, C-shaped (50.67%), V-shaped (26.67%), flat-shaped (15.11%) and L-shaped (7.56%). The angle between the anterior and posterior facets of the flat shape (145.56 ± 9.25)° was the largest and the angle between the anterior and posterior facets of the L shape  (125.07 ± 13.54)° was the smallest(P<0.05); the depth of the notch in the flat shape (3.11 ± 0.83) mm was the smallest and in the L shape (4.47±1.11) mm was the largest(P<0.05);The posterior facet length (13.06 ± 3.56) mm and anterior tibiofibular gap (3.83±1.49) mm on left were larger than on the right side (P<0.05); The posterior facet length (13.36 ± 3.46) mm, fibular notch depth (3.93 ± 1.10) mm and vertical distance of tibiofibular overlap (9.10 ± 2.55) mm larger in men than in women (P<0.05).   Conclusion  In this study, the data related to the inferior tibiofibular syndesmosis notch were measured and divided into four types according to the shape. The flat inferior tibiofibular syndesmosis notch is more likely to have chronic ankle instability, and the fibula is more likely to move forward during anatomical reduction. The inferior tibiofibular syndesmosis of L-shaped and C-shaped notches is more prone to posterior displacement of fibula or poor rotation reduction during anatomical reduction.
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     Finite element analysis of cervical intervertebral discs after removing different ranges of uncinate processes
    YANG Yang SHI Jun LI Kun MA Yuan ZHANG Shao-jie HOU Er-fei WANG Chao-qun CHEN Jie WANG Xing LI Zhi-jun
    2024, 55 (1):  88-97.  doi: 10.16098/j.issn.0529-1356.2024.01.013
    Abstract ( )   PDF (17012KB) ( )  
    Objective  To study the stress change characteristics of the cervical disc after removing different ranges of the uncinate process by establishing a three-dimensional finite element model of the C0-T1 whole cervical spine to provide a theoretical basis for clinical and surgical treatment.    Methods  After a CT scan of one case of the whole cervical spine, we established a three-dimensional model of the normal cervical spine and designed the model to excise 50% and 100% of the uncinate process on the left side and 100% of the uncinate process on both sides at the same time. The four models’ stress changes (stress and displacement) were compared after loading the same torque under six working conditions, forward flexion, back extension, left and right lateral flexion, and left and right rotation.    Results  With the removal of different ranges of the uncinate process, the stress and displacement values of the disc were concentrated at C5/6 and then gradually decreased. Interlateral differences in disc stress values were found only at C5 after removing 50% of the left C5 uncinate process, and interlateral differences were found at all disc stress values after removing 100% of the C5 uncinate process on the left or both sides.    Conclusion  Removal of different ranges of the uncinate process (50% and 100%) can affect cervical spine stability, and the larger the range of resection is the more it affects cervical spine stability. The best clinical principle is to choose the best possible surgical resection in this area that can achieve the least resection and the best therapeutic effect.
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    Risk analysis of re-fracture after percutaneous kyphoplasty in elderly patients with osteoporotic thoracolumbar compression fractures and construction of acolumnar graph prediction model#br#
    SUN Lei WANG Xing-yu XIE Shui-hua
    2024, 55 (1):  98-104.  doi: 10.16098/j.issn.0529-1356.2024.01.014
    Abstract ( )   PDF (2521KB) ( )  
     Objective  To investigate the risk factors for re-fracture after percutaneous kyphoplasty (PKP) in elderly patients with osteoporotic thoracolumbar compression fractures and to construct a line graph prediction model.    Methods  One hundred and eighty-two elderly patients with osteoporotic thoracolumbar compression fractures treated with PKP from January 2016 to November 2019 were selected for the study, and the patients were continuously followed up for 3 years after surgery. Clinical data were collected from both groups; Receiver operating characteristic (ROC) curve analysis was performed on the measures; Logistic regression analysis was performed to determine the independent risk factors affecting postoperative re-fracture in PKP; the R language software 4.0 “rms” package was used to construct a predictive model for the line graph, and the calibration and decision curves were used to internally validate the predictive model for the line graph and for clinical evaluation of predictive performance.   Results  The differences between the two groups were statistically significant (P<0.05) in terms of bone mineral density (BMD), number of injured vertebrae, single-segment cement injection, type of cement distribution, cement leakage, difference in vertebral body height before and after PKP, and change in posterior convexity angle. The area under the curve (AUC) for BMD, number of injured vertebrae, single-segment cement injection volume, cement leakage, pre-and post-PKP vertebral height difference, and posterior convexity change were 0.772, 0.732, 0.722, 0.801, and 0.813, respectively, and the best cutoff values were -3.1, 2, 3.9 ml, 0.4 mm, and 8.7°, respectively. BMD, number of injured vertebrae, single-segment cement injection volume, cement leakage, pre-and post-PKP vertebral height difference, and posterior convexity change were independent risk factors for re-fracture after PKP in elderly patients with osteoporotic thoracolumbar compression fractures. The calibration curve of the column line graph prediction model was close to the original curve and the ideal curve with a C-index of 0.818 (95% CI: 0.762-0.883), and the model fit was good; the threshold value of the column line graph prediction model was >0.22, which could provide a net clinical benefit, and the net clinical benefit was higher than the independent predictors.   Conclusion  BMD, number of injured vertebrae, single-segment cement injection, cement leakage, pre-and post-PKP vertebral height difference, and posterior convexity angle change are independent risk factors affecting the recurrent fracture after PKP in elderly patients with osteoporotic thoracolumbar compression fracture, and this study constructs a column line graph model to predict the recurrent fracture after PKP in elderly patients with osteoporotic thoracolumbar compression fracture as a predictor for clinical. This study provides an important reference for clinical prevention and treatment, and has clinical application value.
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    Histology,Embryology and Developmental Biology
     Retinal microstructure and developmental characteristics in Zebrafish
    FENG Li-ping XU Yi-lin CHEN Xun LIU Da-hai WANG Jun-yong WANG Xiao-ying LIN Jin-xing
    2024, 55 (1):  105-112.  doi: 10.16098/j.issn.0529-1356.2024.01.015
    Abstract ( )   PDF (22473KB) ( )  
    Objective  To study the microscopic structure and morphological characteristics of Zebrafish eyeball and retina at different developmental stages, and to lay a foundation for visual research model.    Methods  Select eight groups of zebrafish at different ages, with six fish in each group, 48 fish in total. Optical microscopy and transmission electron microscopy were used to observe the eyeball structure of Zebrafish at different developmental stages, and the thickness of retinal each layer was measured to analyze the temporal and spatial development pattern. The morphological characteristics of various cells in the retina and the way of nerve connection were observed from the microscopic and ultrastructural aspects, especially the structural differences between rod cells and cone cells.    Results  The retina of Zebrafish can be divided into ten layers including retinal pigment epithelial layer, rod cells and cone cells layer, outer limiting membrane, outer nuclear layer, outer plexiform layer, inner nuclear layer, inner plexiform layer, ganglion cell layer, nerve fiber layer, inner limiting membrane. Rod cells had a smaller nucleus and a higher electron density than cone cells. Photoreceptor terminals were neatly arranged in the outer plexiform layer, forming neural connections with horizontal cells and bipolar cells, and several synaptic ribbons are clearly visible within them. In Zebrafish retina, ganglion cell layer and inner plexiform layer are the earliest developed. With the growth and development of Zebrafish, the thickness of rod cells and cone cells layer and retinal pigment epithelial layer gradually increases, and the retinal structure was basically developed in about 10 weeks.    Conclusion  The retinal structure of Zebrafish is typical, with obvious stratification and highly differentiated nerve cells. There are abundant neural connections in the outer plexiform layer. The ocular development characteristics of Zebrafish are similar to those of most mammals.
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    Technology and Methodology
     Optimization and identification of a low density and high purity method for primary hippocampal neuron culture from fetal rats
    SU Peng WANG Xing-yi LIANG Jing-yan XIONG Tian-qing
    2024, 55 (1):  113-119.  doi: 10.16098/j.issn.0529-1356.2024.01.016
    Abstract ( )   PDF (8358KB) ( )  
    Objective  To establish a low density, high purity and high stability in vitro culture method  of primary hippocampal neurons of fetal rats by co-culturing hippocampal and cortical cells, so as to obtain higher purity and better vitality of primary hippocampal neurons disease.     Methods  The fetal rat hippocampal tissue was isolated from 16-18 days pregnant SD rats, then cut and digested by 0.125% trypsin. The obtained cell suspension was filtered by 200 mesh cell sieve, and then the obtained cell suspensions were then inoculated into the inner layer and outer ring of the culture plate in a surrounding form. They were co-cultured in DMEM/F12 medium containing 10% horse serum. After 4-6 hours of cell adhesion, the culture medium was changed to maintenance medium (Neurobasal+2% B27+0.5 mmol/L glutamine). Then the cell viability was detected with CCK-8 kit and the purity of hippocampal neurons was detected by immunofluorescent staining.   Results  Hippocampal neurons grew well and formed crisscross neural networks after 5 days. And it could survive for 3 weeks. The purity of hippocampal neurons was up to 98%.   Conclusion  The method  of co-culturing hippocampal and cortical cells can obtain high-purity, high activity, high survival rate, and high stability primary hippocampal neurons from fetal rats, which can provide certain experimental conditions for the study of hippocampal neuron related diseases in the nervous system and is worthy of promotion and application.
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     Establishment of a rat model of myocardial hypertrophy by a modified abdominal aortic coarctation method
    HAO Yong-ming GUO Lei ZHOU Cheng-hui PEI Han-jun LI Li ZHAO Zhe
    2024, 55 (1):  120-124.  doi: 10.16098/j.issn.0529-1356.2024.01.017
    Abstract ( )   PDF (5014KB) ( )  
    Objective  To compare effectiveness between the modified and traditional pressure-overload myocardial hypertrophy(POMH) model by abdominal aorta coarctation(AAC) method.    Methods  Totally 45 rats were divided into three groups(n=15 per group), sham group, traditional group, and modified group. In the traditional group, the diameter of the abdominal aorta was narrowed to 0.70 mm through a midline incision for 4 weeks; in the modified group, the diameter of the abdominal aorta was narrowed above the left kidney to 0.45 mm for 1 week, and then the narrowing was lifted postoperatively. The cardiac index, heart weight(HW)/body weight (BW) and left ventricular index, left ventricular weight(LVW)/BW were measured from the heart specimens, and the cross-sectional area of cardiac myocytes, myocardial collagen area, and myocardial collagen area fraction were measured in the pathological sections by HE staining and Masson staining.    Results  Compared with the sham group, the differences in end-systolic interventricular septum thickness(IVSs), left ventricular end-systolic posterior wall thickness (LVPWs), HW/BW, LVW/BW, cardiomyocyte cross-sectional area, myocardial collagen area, myocardial collagen area fraction, and brain natriuretic peptide (BNP) expression levels were statistically significant (P<0.05) in the modified and traditional groups of rats. The differences in these indices were not statistically significant between the modified and traditional groups (P>0.05).    Conclusion  The modified abdominal aortic constriction method  used in this experiment is time-saving, stable, homogeneous and easy to replicate, and is a more ideal approach to establish a rat model of POMH.
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