Table of Content

2023, Volume 54 Issue 2
06 April 2023 Previous Issue    Next Issue
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Review
Anthropology
Cell and Molecules Biology
Anatomy
Histology,Embryology and Developmental Biology
Technology and Methodology
Neurobiology
Anatomy


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Review

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Progress of the talin and the mechanical basis theory of memory CHEN Hui-zhu FANG Xuan TAO Chang-yu ZHANG Wei-guang 2023, 54 (2):  249-254.  doi: 10.16098/j.issn.0529-1356.2023.02.019 Abstract ( )   PDF (1109KB) ( )   Memory loss induced by aging and hypoxia is very common, so exploring the mechanism of memory production, storage and retrieval is of great significance to daily life and clinical work. The storage and retrieval of memory is probably similar to the computer. We summarized the research progress of MeshCODE theory, the mechanical basis of memory. Memory loss in certain diseases (such as Alzheimer’s disease) or pathological conditions (such as aging, lack of oxygen) may be associated with abnormal folding of talin, a mechanosensitive protein. It can dynamically regulate synaptic activity by changing the state of the domain, storing or updating information about small changes in mechanical forces in binary form, and initiating chemical processes such as ligand redistribution in neurons, so that memory is stored in the brain in a binary format, known as the MeshCODE theory.  References | Related Articles | Metrics

Anthropology

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Body composition of Miao nationality in Guizhou and Western Hu’nan LIAN Yi LI Yong-lan YU Hui-xin 2023, 54 (2):  231-237.  doi: 10.16098/j.issn.0529-1356.2023.02.016 Abstract ( )   PDF (3267KB) ( )   Objective  To explore the influence of regional differences on the body composition of the Miao nationality.     Methods  The bioelectrical impedance method  was used to measure 17 body composition indexes of 357 adults of Miao Nationality in Guizhou (162 males and 195 females) and 471 adults of Miao Nationality in Western Hu’nan (210 males and 261 females). The correlation analysis between body composition and age,  One Way ANOVA and principal component analysis were carried out.  Results  The visceral fat grade and trunk fat percentage of Miao men in Guizhou and Miao in Western Hu'nan were positively correlated with age, and total muscle mass and trunk muscle mass were negatively correlated with age. The visceral fat grade and trunk fat rate of Miao women in the two regions were positively correlated with age, and the presumed bone mass and trunk muscle mass were negatively correlated with age. The index values of weight, muscle mass, estimated bone mass, water content, visceral fat grade, limb and trunk muscle mass in Guizhou Miao and Miao men in Western Hu’nan were all larger than women, and the body fat rate, limb and trunk fat mass were all smaller than women. The body fat percentages, limbs and trunk fat percentages of Guizhou Miao men and women were similar to those of Xiangxi Miao, and the muscle mass, limbs and trunk muscle mass were less than that of Xiangxi Miao.   Conclusion  There are  obvious regional differences in muscle mass between the Miao nationality in Guizhou and the Miao nationality in Western Hu’nan.  References | Related Articles | Metrics

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Correlation between sarcopenia,sarcopenic obesity and osteoporosis in adults of Han nationality in Liaoning province LI Jia-yi XI Huan-jiu WEN You-feng GUI Chao ZHANG Xiao-ming ZHANG Hai-long 2023, 54 (2):  238-243.  doi: 10.16098/j.issn.0529-1356.2023.02.017 Abstract ( )   PDF (803KB) ( )   Objective  To understand the prevalence of sarcopenia,sarcopenic obesity and osteoporosis, and to analyze the influence of sarcopenia and sarcopenic obesity on osteoporosis.     Methods  After all the people signed the informed research agreement, the experimental subjects were selected from Han people over 20 years old in Liaoning region, and a total of 1266 cases were included. The distribution of muscle mass, fat mass and bone mineral density in different parts of adults were measured by bioelectrical impedance analyzer (BIA) and bone densitometer, and the correlation between sarcopenia and sarcopenic obesity and osteoporosis in adults was studied.     Results  With the increase of age, the muscle mass of limbs, trunk, total muscle mass and body weight of the Han adults in Liaoning showed a trend of increasing at first and then decreasing. There was no significant difference in the prevalence of sarcopenia and sarcopenic obesity between men and women, but there was a significant difference in the prevalence of osteoporosis between men and women. The prevalence of all three groups reached the peak in the age group above 60, and the difference was statistically significant.The risk factor for osteoporosis was sarcopenia and sarcopenic obesity.     Conclusion  Among adults of Han nationality in Liaoning, the prevalence of sarcopenic obesity,sarcopenia and osteoporosis is significantly different in age. Bone condition is affected by fat mass and muscle mass. References | Related Articles | Metrics

Cell and Molecules Biology

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Mechanism of microRNA-30d-5p regulating osteogenic differentiation of bone marrow stromal cells through glucose-regulated 78 LI Guang LI Jie ZHANG Ping 2023, 54 (2):  195-201.  doi: 10.16098/j.issn.0529-1356.2023.02.010 Abstract ( )   PDF (10599KB) ( )   Objective　To investigate the effect of miR-30d-5p on osteogenic differentiation and apoptosis of bone marrow stromal cells and its mechanism. Methods　Bone marrow stromal cells were divided into miR-30d-5p overexpression negative control group, miR-30d-5p overexpression group, miR-30d-5p inhibition negative control group and miR-30d-5p inhibition group. Alkaline phosphatase (ALP) staining was used to identify osteogenesis, alizarin red staining was used to detect calcium nodules precipitation, and TUNEL was used to detect apoptosis. mRNA and protein expression levels were detected by qPCR and Western blot, and the potential binding sites of miR-30d-5p were predicted by the bioinformatics analysis website Targetscan7.1. Results　After miR-30d-5p overexpression, osteogenic differentiation ability, and mineralization ability of the cells decreased (P<0.05), while apoptosis level increased (P<0.05). The expression of glucoregulatory protein 78 (GRP78) and osteogenic specific transcription factor RUNX2 was significantly decreased (P<0.05). However, miR-30d-5p inhibitor-treated the cells with increased osteogenic differentiation and mineralization ability (P<0.05), and apoptosis level decreased (P<0.05). GRP78 and RUNX2 protein levels increased (P<0.05). The miR-30d-5p binding site is located at 142-148bp of the 3'UTR of the GRP78 gene. Conclusion　MiR-30d-5p inhibits osteogenic differentiation and promotes apoptosis of bone marrow stromal cells by down-regulating the expression of GRP78 protein. References | Related Articles | Metrics

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Expression change and role of sex determining region box transcription factor 2 mRNA and the non-coding RNA in the hepatocyte stem changes during the rat  liver regeneration WANG Zi-hui GUO Jian-lin ZANG Xia-yan XUE Qi-jie LIN Kai-lin ZHANG Chun-bo HAN Lu LIN Jun-tang XU Cun-shuan 2023, 54 (2):  202-207.  doi: 10.16098/j.issn.0529-1356.2023.02.011 Abstract ( )   PDF (4578KB) ( )   Objective  To explore the role pathway and pattern of the sex determining region box transcription factor 2 (SOX2) and its mRNA interaction with microRNA(miRNAs, miR) and circular RNA(circRNA) at 0 hour and 2 hours in the rat liver regeneration.     Methods  The rat 2/3 hepatectomy (partial hepatectomy, PH) model was prepared as described by Higgins, the hepatocytes were isolated according to the method  of Smedsrod et al, the expression changes of mRNA, miRNA and circRNA ［together named as competing endogenous RNAs(ceRNA)］ were detected by the large-scale quantitative detection technology, the interaction network of ceRNA was constructed by Cytoscape 3.2 software, and their correlation in expression and role were analyzed by ceRNA comprehensive analysis.    Results  It was found that at the 0 hour and 2 hours after PH, the ratio value of SOX2 mRNA shows 1.00±0.09 and 2.15±0.48, miR-3558-3p displays 4.53±0.10 and 0.81±0.16, circRNA_18404 shows 1.24±0.04 and 11.10±0.57, circRNA_18045 displays 1.97±0.47 and 4.44±0.23. At the same time, the eight kinds of cell dedifferentiation-related genes AT-rich interaction domain 5A (ARID5A), activating transcription factor 3 (ATF3), BTG anti-proliferation factor 2 (BTG2), etc, which are prometed in expression by SOX2, were down-regulated at 0 h after PH, but the cell differentiation-related genes interferon regulatory factor 6 (IRF6) and somatostatin (SST), which are inhibited in expression by SOX2, were up-regulated at 0 hour after PH. On the other hand, the eight kinds of cell dedifferentiation-related genes ARID5A, ATF3, BTG2, etc, which are promoted in expression by SOX2, were up-regulated at 2 hours after PH, but the cell differentiation-related gene SST, which is inhibited in expression by SOX2, was down-regulated, and IRF6 had no meaningful changes in expression at 2 hours after PH.    Conclusion  The correlation in expression and role of the miRNA, which are inhibited by circRNA, SOX2, its mRNA is inhibited by miRNA, and the cell stem-related genes, which are regulated by SOX2, are helpful for the hepatocyte to be in differentiation state at 0 hour after PH and to be in stem state at 2 hours after PH.  References | Related Articles | Metrics

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Association between hydroxysteroid dehydrogenases gene family single-nucleotide polymorphisms and digit ratio LI Fan MA Zhan-bing ZHANG Jing YANG Meng-yi WANG Yuan MA Qian LU Hong HUO Zheng-hao DANG Jie 2023, 54 (2):  208-215.  doi: 10.16098/j.issn.0529-1356.2023.02.012 Abstract ( )   PDF (6064KB) ( )   Objective  investigate the association between 8 single-nucleotide polymorphisms (SNPs) of hydroxysteroid dehydrogenase (HSD) gene family and human digit ratio (2D ∶4D).     Methods  Randomly selected 808 college students (400 males and 408 females) as subjects, the digit ratio of left and right fingers were measured and calculated using computer image software. Eight SNPs (rs1000283, rs2236903, rs5479, rs56303414, rs676387, rs4445895, rs2066474, rs8190478) in HSD11B and HSD17B gene families were genotyped by multiplex PCR. The association between 2D∶4D and different genotypes was analyzed by One-Way ANOVA.    Results  Female left hand(L)2D ∶3D, L2D ∶4D, L3D ∶4D, right hand(R)2D ∶4D, R2D ∶5D were significantly higher than male (P<0.05). The genotype frequency of rs2236903 (HSD11B1) was significantly different between male and female (P <0.05). A good linkage disequilibrium was found between rs1000283and rs2236903 of HSD11B1 gene, rs5479 and rs56303414 of HSD11B2 gene, but there was no statistical difference in frequency between male and female students(P>0.05). The genotypes frequency of the 8 SNPs were not significantly associated with digit ratio (2D ∶4D) in both males and females (P >0.05).     Conclusion  There are significant gender differences in digit ratio in Ningxia Han college students, but there is no correlation between digit ratio and 8 SNPs in HSD11B and HSD17B gene families, suggesting that HSD11B and HSD17B gene families may have nothing to do with the formation of human digit ratio.  References | Related Articles | Metrics

Anatomy

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Existence and morphology features of the myodural bridge in JapaLura Splendida  BAI Xue KANG Hao-nan DOU Ya-ru TANG Wei ZHENG Nan SUI Hong-jin 2023, 54 (2):  220-225.  doi: 10.16098/j.issn.0529-1356.2023.02.014 Abstract ( )   PDF (5887KB) ( )   Objective  The dense fibrous connective tissue that connects sub-occipital muscles which consist of the rectus capitis posterior minor muscle (RCPmi), rectus capitis posterior major muscle (RCPma), obliquus capitis inferior muscle (OCI) and nuchal ligament (NL) to the spinal dura mater (SDM), is described as the myodural bridge (MDB) in humans. The MDB is perceived as an essential anatomical structure and has been a subject of interest for clinicians. Studies have revealed that MDB may be related to the dynamic circulation of the cerebrospinal fluid (CSF) and a chronic cervicogenic headache. To date, the MDB is identified as a universal, existing structure in mammals and it exists in other vertebrates as well, such as Gallus domesticus and Rock pigeons in Avifauna, Siamese crocodile and Trachemys scripta elegans in Reptile. The current study is to further analyze different structures features of the MDB in sundry classes and provide the anatomical basis for functional studis  The JapaLura Splendida is the most common species in Lacertiformes, Reptilia. So we chose it as the experimental object to supply the morphological study of the MDB in Reptilia.      Methods The study  was based on gross anatomical dissection, thick sheet section, histological staining to observe the structural characteristics of the post-occipital area of twenty JapaLura Splendidas and the existence of the MDB.     Results  The deep post-occipital muscles were composed of the rectus capitis dorsal muscle (RCD) and the obliquus capital posterior (OCP) muscle. The RCD was merged by the rectus capitis dorsal major muscle (RCDma), the rectus capitis dorsal minor muscle (RCDmi) and the obliquus capitis anterior muscle (OCA). In the atlanto-occipital space, the dense fibrous bundles were found to originate from the ventral aspect of the RCD and run ventral, closely inserting into the SDM. In the atlanto-axial space, the dense fibrous bundles were found to originate from the ventral aspect of the OCP and run ventral, closely contacted with the SDM. These dense fibrous bundles were the collagen type I fibers with strong double refraction.     Conclusion  The result  of this study indicates that the MDB is located between the post-occipital muscles and the SDM in JapaLura Splendida. The MDB of Japalura splendida may be related to the activities of the head and neck, and exert a physiological function similar to the MDB in humans. References | Related Articles | Metrics

Histology,Embryology and Developmental Biology

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Monitoring second polar body exclusion by time-lapse in predicting fertilization and embryo development efficiency of intracytoplasmic sperm injection Lin-Tao XUE Shikai Wang Xianbao Mao Zhengda Li Xiaohui Zhang Pinpin Wei 2023, 54 (2):  226-230.  doi: 10.16098/j.issn.0529-1356.2023.02.015 Abstract ( )   PDF (2335KB) ( )   Objective  To explore the clinical value of second polar body (Pb2) exclusion monitoring by time-lapse in predicting the fertilization and embryo development efficiency for intracytoplasmic sperm injection (ICSI).     Methods  A retrospective research was performed on 278 patients treated with ICSI, the clinical data and Time-lapse monitoring embryo culture data were collected and analyzed, to explore the exclusion of Pb2 after ICSI and the relationship between the specific exclusion time and the outcome of fertilization and embryo development.     Results  The average time of Pb2 exclusion after ICSI was (3.03 ± 1.21) hours; The fertilization rate, 2 pronucleus(PN) fertilization rate and 5 days(D5) blastocyst formation rate in the Pb2 exclusion group were significantly higher than those in the without Pb2 exclusion group (99.95% vs 1.75%, P<0.001; 97.18% vs 0.66%, P<0.001; 60.50% vs 16.67%, P<0.05); The 2PN fertilization rate in Pb2 exclusion time >3-4 hours group was significantly higher than that in 0-2 hours group and >5 hours group (98.80% vs 93.81%, P<0.05; 98.80% vs 95.40%, P<0.05); The exclusion time of Pb2 was significantly correlated with the average number of blastomeres in D3 embryos (P<0.001). The D5 blastocyst formation rate of 3-4 hours group was significantly lower than that of >2-3 hours group (56.23% vs 67.23%, P<0.05), >4-5 hours group was significantly lower than 0-2 hours group and >2-3 hours group (46.61% vs 62.30%,  P<0.05; 46.61% vs 67.23%,  P<0.05), and D5 blastocyst formation rate of >5 hours group was 7.14%, which were significantly lower than that of the other four groups ( P<0.05). The formation rate of D5 high-quality blastocysts in 3-4 hours group was significantly lower than 0-2 hours group and > 2-3 hours group (9.92% vs 16.39%,  P<0.05; 9.92% vs 20.72%,  P<0.05), and D5 high-quality blastocysts formation rate in > 4-5 hours group was significantly lower than that in > 2-3 hours group (11.02% vs 20.72%,  P<0.05).     Conclusion  Monitoring Pb2 exclusion by Time-lapse can accurately predict fertilization outcome. The time of Pb2 exclusion is significantly correlated with embryo development potential. It is a valuable morphological index to predict fertilization and embryo development outcome in ICSI. References | Related Articles | Metrics

Technology and Methodology

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Design and validation of a surgical guide for the retrieval of foreign body instruments in jaw MAO Meng-yun WANG Cheng-ze ZHU Hai-hua FAN Ming-xing WU Xiao-fei ZHU Fu-dong 2023, 54 (2):  244-248.  doi: 10.16098/j.issn.0529-1356.2023.02.018 Abstract ( )   PDF (9669KB) ( )   Objective  To design and validate a novel surgical guide for retrieval of foreign body in jaw.     Methods Firstly, a surgical guide based on cone beam computed tomography and trephine technique was designed to remove broken dental instrument fragments. Its feasibility and accuracy were assessed by removing broken dental instrument in goat mandible, and then it was successfully applied in clinical cases. The linear and angular discrepancies between actual and planned columnar bone with imaginary fragment was analyzed. The euclidean distance was measured at the hex and apex of the columnar bone and the angle of axis deviation was also calculated. We obtained seven parameters (cdh, cda, hdh, hda, vdh, vda, and ad) to describe deviations.    Results  Mean central deviation at the hex and apex was (0.51 ± 0.14) mm and (0.62 ± 0.19) mm, respectively. Accompanying mean values were as follow: horizontal deviation at the hex was (0.48 ± 0.16) mm, horizontal deviation at the apex was (0.52 ± 0.22) mm, vertical deviation at the hex was (0.17 ± 0.09) mm, vertical deviation at the apex was (0.29 ± 0.13) mm, and angular deviation of (5.38 ± 3.43) degrees. In a clinical case, the guide successfully located and removed the fracture file.    Conclusion  This study reveals that this kind novel surgical guide could aid to locate and remove the foreign body in jaw.  References | Related Articles | Metrics

Neurobiology

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Alterations in dendritic length and branch and K+ -Cl - cotransporter 2 expression of the olfactory bulb in young APP/PS1 mice CHEN Bo-yu JIANG Jin-xiang ZHANG Jia-wei YANG Li 2023, 54 (2):  127-133.  doi: 10.16098/j.issn.0529-1356.2023.02.001 Abstract ( )   PDF (1793KB) ( )   Objective  To study the morphology of olfactory bulb(OB) neurons and the change of related proteins, and explore the causes of olfactory dysfuction in Alzheimer’s disease(AD).    Methods  Golgi-Cox staining technique was used to evaluate the morphological changes of neurons in the OB and anterior piriform cortex (aPC) of APP/PS1 AD model mice. The morphology of neurons was determined by Sholl analysis. Western blotting was used to evaluate the levels of protein expression.    Results  The results of Golgi-Cox showed that the dendrite length and branch number reduced significantly in the OB neurons of 3-5-month-old APP/PS1 mice, an age that the mice did not show the pathological characteristics and cognitive impairment of AD. Western blotting analysis showed that levels of potassium chloride cotransporter 2（KCC2）, a potassium chloride transporter crucial for neuronal morphology and synaptic function,   decreased significantly in the OB of 3-5-month-old APP/PS1 mice.    Conclusion  Abnormal neuronal morphology and KCC2 signal might be the basis of early olfactory dysfunction in AD. Thus, maintaining normal KCC2 signal may be one of the keys to intervene the olfactory abnormalities in the early stage of AD.   References | Related Articles | Metrics

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Effects of chronic restraint stress on the expression of N6-methyladenosine and related enzymes in the hippocampus of mice ZHANG Yuan SHI Meng-meng HE Jia-bei WU Qiong ZHONG Xiao-lin XIANG Yu-yan XU Yang HE Jie 2023, 54 (2):  142-148.  doi: 10.16098/j.issn.0529-1356.2023.02.003 Abstract ( )   PDF (2364KB) ( )   Objective  To investigate the effect of chronic restraint stress on the expression of N6-methyladenosine (m6A）and related enzymes in the hippocampus of mice.     Methods Twenty C57BL/6J male mice were randomly divided into control group and chronic restraint stress (CRS) group, the model group was given for 3 weeks chronic restraint stress to establish a mouse anxiety model. Open field test and elevated plus maze test were used to detect anxiety-like behavior; Immunohistochemistry and m6A RNA methylation assay were used to detect the expression changes of mouse hippocampal m6A; Western blotting and Real-time PCR were used to analyze hippocampal m6A related enzymes expression.     Results  1.The behavioral results showed that, compared with the control group, the CRS group showed significantly reduced time spent in the center of the open field(P<0.01), the CRS group showed significantly reduced exploration time in the open arm of elevated plus maze (P<0.0001); 2. Immunohistochemical results showed that, compared with the control group, the hippocampal m6A content in the CRS group reduced significantly (P<0.001); The results of the m6A RNA methylation assay showed that, compared with the control group, the CRS group showed significantly reduced amount of hippocampal m6A(P<0.05); 3. Real-time PCR results showed that the expression of hippocampal demethylase anaplastic lymphoma kinase B(AlkB) homolog 5(ALKBH5) (P<0.001) and fat mass and obestity associated protein(FTO) (P<0.05) in the CRS group significantly up-regulated, the expression of methylase Wilms’tumour 1-associating protein(WTAP) (P<0.05) was significantly down-regulated compared with the control group; The expression of m6A methylation binding protein YTH domaincontaining family protein 3(YTHDF3) (P<0.05) and YTH domaincontaining protein 2(YTHDC2)(P<0.01) was significantly up-regulated. Western blotting  result  showed that, compared with the control group, the mouse hippocampal demethylase ALKBH5 (P<0.05) and FTO (P<0.05) expression in the CRS group significantly up-regulated, the expression of WTAP (P<0.01) was significantly down-regulated; m6A methylation binding protein YTHDF3 (P<0.01) and YTHDC2 (P<0.05) were significantly up-regulated.     Conclusion In the anxiety model induced by chronic restraint stress, the expression of m6A in the hippocampus of mice is down-regulated. The mechanism may be related to the up-regulation of the m6A demethylase ALKBH5 and FTO or the down-regulation of the methylase WTAP.  References | Related Articles | Metrics

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Effects of transient receptor potential vanilloid 4 inhibitor HC067047 on anxiety-like behavior in mice induced by lipopolysaccharide GUO Dong-ming ZHANG Jing-wen HUANG Xin-lei XU Yang XIONG Tian-qing CAO Wen-yu LIANG Jing-yan 2023, 54 (2):  149-155.  doi: 10.16098/j.issn.0529-1356.2023.02.004 Abstract ( )   PDF (3645KB) ( )   Objective  To investigate the effect of transient receptor potential vanilloid 4 (TRPV4）inhibitor HC067047 on anxiety-like behavior in mice induced by lipopolysaccharide (LPS).    Methods  Totally 48 male C57BL/6 mice were randomly divided into control group (NS), model group (LPS) and drug intervention group (HC+LPS). Anxiety mouse model was established by intraperitoneal injection of 0.83 mg/kg LPS. The HC+LPS group was intraperitoneally injected with HC067047 (10 mg/kg) 30 minutes before LPS injection, and the NS group and LPS group were injected with equal volume of normal saline. Open field test and social interaction experiments were used to detect anxiety-like behaviors in each group of mice; Immunohistochemical chemistry and Western blotting were used to detect the expression of TRPV4, inducible nitric oxide synthase (iNOS), neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) in the hippocampus.    Results  Immunohistochemical and Western blotting experiments showed that, compared with the NS group, the expression of TRPV4 in the hippocampus of the LPS group was significantly up-regulated (P＜0.0001); In the open field test, compared with the NS group, the total distance (P<0.0001), the distance in the central area (P<0.01) and the time of in the central area mice in the LPS group  reduced significantly (P<0.01). HC067047 intervention reversed the activities of LPS model mice total distance (P＜0.05), the distance of activities in the central area (P＜0.001) and the time of in the central area (P＜0.01); In the social interaction test, compared with the NS group, the interaction time the unfamiliar mice reduced significantly in LPS group (P<0.01), which was reversed by HC067047 treatment (P< 0.01); Western blotting detection revealed that the expression of hippocampal iNOS (P＜0.05), nNOS (P＜0.001), and eNOS (P＜0.001) in the LPS group were significantly higher than the NS group, which reduced remarkably by HC067047 treatment (iNOS  P<0.05, nNOS  P<0.01 and eNOS  P<0.01).     Conclusionn Inhibiting the expression of TRPV4 can improve the anxietylike behavior, and this process may be related to anti-oxidative stress.  References | Related Articles | Metrics

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Effect of altitude hypoxia on blood-brain barrier after subarachnoid hemorrhage based on phosphatidylinositol 3-kinase/protein kinase B/nuclear factor  κBpathwayin rats WEI Yan-na WANG Feng-cun MA Xiang-lian SUO Tian-sha CHEN Sheng WANG Lan-gui SUN Juan ZHAO Xiu-li 2023, 54 (2):  156-164.  doi: 10.16098/j.issn.0529-1356.2023.02.005 Abstract ( )   PDF (5680KB) ( )   Objective  To investigate the effect of plateau hypoxia on the blood-brain barrier after subarachnoid hemorrhage (SAH) in rats.   Methods  Adult male SD rats (n=78) were randomly divided into 4 groups: sham group (sham), SAH model group (SAH), plateau hypoxia sham group (Hp sham) and plateau hypoxia SAH model group (Hp SAH). The rat model of plateau hypoxia was established through low-pressure simulation chamber (altitude 5000 m), and the SAH model was established by endovascular perforation method. At 24 hours after SAH, neurobehavior score and SAH grade were assessed. The morphological changes of neurons and apoptosis of nerve cells in the CA1 region of hippocampal were observed by the staining of Nissl and TUNEL. The expression of phosphorylated PI3K (p-PI3K), PI3K, phosphorylated Akt (p-Akt), Akt, phosphorylated nuclear factor κB(p-NF-κB), NF-κB, matrix metalloproteinase-9 (MMP-9), occludin and claudin-5 in hippocampal were detected by the method  of Western blotting. The expression of occludin and claudin-5 proteins in the CA1 region of hippocampal were observed by immunofluorescent staining.    Results  At 24 hours after SAH, the neurobehavior score decreased significantly and SAH grade increased significantly in the SAH and Hp SAH group (P <0.05). Neurobehavior score decreased significantly in the Hp SAH group compared with the SAH group (P< 0.05). In the SAH group, neurons in the CA1 region of hippocampus were atrophied and deformed, the arrangement were disordered, the number of neurons decreased significantly, and the apoptosis of nerve cells increased significantly(P< 0.05). Plateau hypoxia could aggravate the morphological damage of neurons and apoptosis of nerve cells. The expression of p-PI3K/PI3K, p-Akt/Akt, occludin and claudin-5 proteins decreased significantly, while the expression of p-NF-κB/NF-κB and MMP-9 proteins increased significantly in the SAH and Hp SAH group (P< 0.05). The expression of p-PI3K/PI3K and MMP-9 proteins increased significantly in Hp SAH group compared with the SAH group. The expression of claudin-5 protein increased significantly in Hp sham group compared with the sham group (P<0.05). Immunofluorescent staining showed that the expression of occludin and claudin-5 proteins in the CA1 region of hippocampus decreased in the SAH group. Plateau hypoxia could further decreased the expression of occludin and claudin-5 proteins. Conclusion  Plateau hypoxia aggravates blood-brain barrier disruption after subarachnoid hemorrhage in rats through inhibiting PI3K/Akt/NF-κB pathway.  References | Related Articles | Metrics

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Impaired cholesterol metabolism affecting subventricular zone neurogenesis in ob/ob mice LI Rui-ting LEI Ling-feng YANG Na XIANG Zhi-yu LI Ze-kai LU Li 2023, 54 (2):  165-174.  doi: 10.16098/j.issn.0529-1356.2023.02.006 Abstract ( )   PDF (8887KB) ( )   Objective  To investigate the effect of cholesterol on the proliferation and differentiation of neural stem cells (NSCs) in ob/ob obese mice, and to explore the possible mechanism of central nervous systym dysfunction caused by obesity.     Methods  Selected 6 4-month-old ob/ob and wild type (WT) mice, and cell proliferation antigen (Ki67) and doublecortin (DCX) immunofluorescenct staining were used to detect ob/ob mice lateral ventricle subventricular zone (SVZ) neurogenesis level. Cultured SVZ NSCs isolated from 18 4-month-old ob/ob and WT mice, and BrdU incorporation experiment and β-Ⅲ-tubulin (Tuj1) immunofluorescent staining were employed to detect the self-renewal and differentiation ability of NSCs. Matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI- MS）was used to detect the lipid distribution in 4-month-old ob/ob and WT mice brain tissues, and measure the changes of cholesterol(ST) content and the expression genes related to cholesterol synthesis. Cultured 15 WT postnatal day 0(P0) mouse SVZ NSCs in vitro and electrotransfected with the small interfering RNA(siRNA) sequence of cholesterol synthesis rate-limiting enzyme 3-hydroxy-3-methyl-glutaryl coenzyme A reductase（Hmgcr）verified the knockdown efficiency, to detecte the effect of Hmgcr gene knockdown on NSCs by BrdU incorporation experiment and Tuj1 immunofluorescent staining.     Results  Compared with the WT mice, the number of Ki67+ and DCX+ cells in the SVZ tissue of ob/ob mice decreased significantly (P<0.05); The result  of in vitro experiments showed that the BrdU positive rate (P<0.05) and Tuj1 positive rate (P<0.05) of ob/ob mice NSCs were also significantly lower than that of WT mice NSCs. MALDI-MS test result  showed that the lipid distribution of ST (26∶0) and ST (27∶3) reduced significantly on SVZ of ob/ob mice. Meanwhile, Kit assay result  showed that the cholesterol content of ob/ob mice SVZ tissue and NSCs both reduced significantly (P<0.05). In addition, the Real-time PCR result  showed that the mRNA expression levels of cholesterol synthesis-related genes Hmgcr, Lanosterol 14-alpha demethylase（Cyp51）, 3-beta-hydroxysteroid-Δ8, Δ7-isomerase（Ebp）and squalene synthase（Fdft1）in the SVZ tissue and NSCs of ob/ob mice were both significantly lower than those in WT mice (P<0.05), suggesting that ob/ob mice SVZ cholesterol synthesis disordered. Real-time PCR result  showed that the interference efficiency of siRNA was 55.27%±8.768% (P<0.05), and the result  of Western blotting showed that the interference efficiency of siRNA was 32.69%±8.056%(P<0.05). The result  of BrdU incorporation experiment indicated that the positive rate of BrdU in the Hmgcr-siRNA group was significantly lower than that in the control group (P<0.05). After differentiation induced with 1% FBS, the positive rate of Tuj1 in the Hmgcr-siRNA group also decreased significantly compared with the control group (P<0.05), suggesting that impaired cholesterol synthesis inhibited the self-renewal and differentiation ability of neural stem cells.      Conclusion  Decreased cholesterol levels in the ob/ob mice brain can affect the proliferation and differentiation of SVZ NSCs.  References | Related Articles | Metrics

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Polyinosinic-polycytidylic acid affecting the expressions of Bcl-2 and Bax after cerebral ischemia-reperfusion in hyperlipidemic rats and its neuroprotective effect GAO Sai-hong ZHANG Xiao-liang YANG Ying-chun QIAO Hai-bing 2023, 54 (2):  175-180.  doi: 10.16098/j.issn.0529-1356.2023.02.007 Abstract ( )   PDF (6795KB) ( )   Objective  To observe the effect of polyinosinic-polycytidylic acid (Poly-IC) treatment on the expressions of Bcl-2 and Bax after cerebral ischemia-reperfusion （I/R）injury in hyperlipidemia rats, and to detect the cerebral infarction, blood-brain barrier permeability and behavioral injury symptoms, to explore the neuroprotective effect of Poly-IC treatment on cerebral I/R injury in hyperlipidemia rats.    Methods  Hyperlipidemia rats were randomly divided into cerebral I/R group, Poly-IC pretreatment group, Poly-IC post-treatment group and sham operation group, 20 rats in each group. Neurobehavioral performance of rats in each group was recorded according to neurobehavioral score of 0-4 points. Blood-brain barrier permeability of rats in each group was detected by Evans blue staining. TTC staining was used to observe the cerebral infarction in each group. Apoptotic cells in the cerebral cortex of rats in each group was observed by TUNEL staining. The relative expression levels of Bcl-2 and Bax were determined by Western blotting.    Results  Compared with the sham group, the symptoms of neurobehavioral damage in the I/R group were serious and the score increased significantly(P<0.05). The scores of Poly-IC pretreatment and post-treatment groups were significantly lower than that of I/R group(P<0.05). Evans blue staining result  showed that the blood-brain barrier permeability of the I/R group was significantly higher than that of the sham group (P<0.05), and Poly-IC pretreatment or post-treatment could significantly reduce the blood-brain barrier permeability(P<0.05). No infarct was observed in the sham group with uniform red staining, while white infarct was observed in the brain tissue of the I/R group. Compared with the I/R group, the volume of infarct in both Poly-IC pretreatment and post-treatment groups reduced significantly (P<0.05). The apoptosis index in cerebral cortex of rats in I/R group was significantly higher than that in sham group (P<0.05), while the apoptosis index in Poly-IC pretreatment or post-treatment group was significantly lower than that in I/R group(P<0.05). The result  of Western blotting showed that, compared with the sham group, the expression of Bax in the I/R group was significantly increased(P<0.05), the expression of Bcl-2 was significantly decreased(P<0.05). Compared with the I/R group, the expression of Bax in the Poly-IC pretreatment or post-treatment group reduced significantly(P<0.05), the expression of Bcl-2 increased significantly(P<0.05).     Conclusion  Poly-IC pretreatment or post-treatment can improve the symptoms of neurobehavioral injury, reduce the damage of blood-brain barrier, reduce the volume of cerebral infarction, decrease the apoptosis index of nerve cells, play a neuroprotective effect on cerebral ischemia reperfusion injury in rats with hyperlipidemia, and this protective effect may be related to the change of Bcl-2 and Bax expression levels. References | Related Articles | Metrics

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Dexmedetomidine improving complete Freund’s adjuvant-induced anxiety-like and depression-like behaviour by promoting the expression of brain-derived  neurotrophic factor-tyrosin kinase receptor B in mice hippocampus WANG Ya-juan YANG Bin ZENG Xue-qing LIANG Yue LIU Zhi-wen CAO Wen-yu ZHONG Xiao-lin 2023, 54 (2):  181-187.  doi: 10.16098/j.issn.0529-1356.2023.02.008 Abstract ( )   PDF (3001KB) ( )   Objective  To study the effect of dexmedetomidine (DEX), an α2- adrenoceptor agonist, on the pain-related anxiety-like and depression-like behaviour induced by complete Freund’s adjuvant (CFA) injection and its possible regulatory mechanism.  Methods  Thirty-six ICR female mice were randomly divided into normal saline (NS) group, CFA group and DEX+CFA group, n=12 for each group. Chronic inflammatory pain model was established by subcutaneous injection of 10 μl CFA into the right hind limb of mice. DEX+CFA group mice were injected intraperitoneally with 0.025 mg/kg DEX 30 minutes before nociceptive behavior test, and once a day for 7 days. Von-frey fiber was used to evaluate the threshold of mechanical pain in mice, n=12 for each group. The anxiety-like behavior of mice were detected by open field test, n=12 for each group. Sucrose preference, tail suspension test and forced swimming test were used to detected the depression-like behavior of mice, n=12 for each group. The expression of adrenergic receptor β2 (ADRB2), Brain-derived neurotrophic factor (BDNF), tyrosine kinase B receptor (TrkB), and glutamate receptors 1(GluR1) and GluR2 were detected by Western blotting, n=8 for each group. Immunohistochemical staining was used to detect the expression of recombinant doublecortin(DCX), which is a marker of newborn neurons in the hippocampus, n=4 for each group.    Results  Compared with the NS group, the mechanical threshold of mice on the 1st, 3rd and 7th day after CFA injection decreased significantly (P<0.05); But there was no significant difference between DEX+CFA group and CFA group (P>0.05). Compared with the NS group, the time spent in the inner ares (P<0.01), number of entering the central grid area (P<0.01) and distance travelled in the inner area (P<0.01) of CFA group mice reduced significantly, while the time (P<0.01), numbers (P<0.05) and distance（P<0.05）of DEX+CFA group mice entering the central grid area enhanced significantly. The result  of depression-like behavior tests showed that the sucrose preference percentage (P<0.05) reduced  significantly in CFA group when compared with NS group, and the immobility time increased significantly in tail suspension test (P<0.01) and forced swimming test (P<0.001) in CFA mice when compared with NS group, while DEX intervention could significantly increase the sucrose preference scores (P<0.05) and decreased the immobility time in tail suspension test (P<0.05) and forced swimming test (P<0.05). The result  of Western blotting showed that compared with the NS group, the levels of ADRB2 (P<0.0010), BDNF (P<0.001), TrkB (P<0.01), GluR1 (P<0.001) and GluR2 (P<0.001) in the hippocampus of CFA group were significantly decreased, while DEX intervention could significantly increase the expression of ADRB2 (P<0.05), BDNF (P<0.001), TrkB (P<0.001), GluR1 (P<0.001) and GluR2 (P<0.001). Immunohistochemical result  showed that compared with the NS group, the average absorbance(AA) of DCX decreased significantly in hippocampus of CFA group (P<0.05), but increased significantly in DEX+CFA group (P<0.05).    Conclusion  Dexmedetomidine may promote hippocampal neurogenesis through upregulated the expression of BDNF-TrkB, thus improving CFA-induced anxiety-like and depression-like behaviors in mice. References | Related Articles | Metrics

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Expression and localization of neuroglobin in the brain tissue of Saiga antelope LIU Xia ZHENG Li-ping DU Xiao-hua WANG Yu-jiao WU Ya-juan 2023, 54 (2):  188-194.  doi: 10.16098/j.issn.0529-1356.2023.02.009 Abstract ( )   PDF (17141KB) ( )   Objective  Saiga antelope is a small population inhabiting in desert and semi desert areas of national and world endangered protected animals, its wild population is extremely rare. In order to explore the correlation between hypoxic tolerance and neuroglobin (NGB) in Saiga antelope. A female Saiga antelope died of dystocia was used as the experimental animal, and the tissue samples were sampled repeatedly for 3 times to study the distribution and expression of NGB in brain of Saiga antelope in the process of adapting to hypoxia.     Methods  The distribution and expression of NGB in the parietal lobe, frontal lobe, temporal lobe, occipital lobe, hypothalamus, hippocampus, pear like leaf, cingulate gyrus, striatum and thalamus of Saiga antelope were detected by immunohistochemistry(IHC) and Real-time PCR.      Results  The result  of IHC showed that NGB was positive in all parts of  Saiga antelope brain, and the cells that had positive reactions in the parietal, frontal, temporal and occipital lobes of the cerebral cortex were mostly granular cells and martinotti cells. NGB was found in the granular cell layer, pyramidal cell layer and molecular cell layer in hippocampus, and the positive staining of pyramidal cell layer was the strongest. NGB positive expression in Pear like leaves and hypothalamus mainly occured in multi-type cells. NGB was expressed in the granulocytes and glial cells of cingulate gyrus, mainly in the granular cells. The positive expression of NGB in striatum was mainly located in granular cells, the positive expression of NGB in thalamus could be seen in the polymorphosis and glial cells, and the positive substance of the multi-type cells was obviously colored. The result of Real-time PCR showed that NGB was expressed in different regions of Saiga antelope brain, the highest expression in the frontal lobe of the cerebral cortex, the second in the parietal lobe, and the expression was significantly higher than that in the rest of the brain tissue (P<0.05). The expression of hippocampus, hypothalamus and thalamus was significantly higher than that in the rest (P<0.05). The expression of pear like leaves was significantly higher than that of temporal lobe, occipital lobe, cingulate gyrus and striatum (P<0.05). The expression of temporal and occipital lobe was significantly higher than that of cingulate gyrus and striatum (P<0.05). The expression of the cingulate gyrus and striatum was the lowest (P>0.05).      Conclusion  The expression of NGB in different regions of Saiga antelope has some selective differences in the long-term adaptation to hypoxia environment. The frontal and parietal lobes have the highest tolerance to hypoxia, followed by hippocampus, and the striatum is the weakest, which may be related to the specific functions of different regions of brain tissue, but the specific mechanism remains to be further explored. References | Related Articles | Metrics

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Spatial and temporal expression patterns of Atoh1 during mice cerebellar development XUE Sai-sai CHEN Li-ping LI Ying WU Yan WU Hai-tao 2023, 54 (2):  134-141.  doi: 10.16098/j.issn.0529-1356.2023.02.002 Abstract ( )   PDF (6082KB) ( )   Objective  To investigated the spatial and temporal expression of Atoh1 mRNA and protein level during the cerebellar development.   Methods  The frozen cerebellum sections of mice at different developmental stages were used to analyze the temporal and spatial expression patterns of Atoh1 mRNA by RNA scope technique, n=3 for each group. Meanwhile, two reporter mice with different genetic modification method were utilized to analyze the   spatialand temporal expression of Atoh1 at the protein level by immunofluorescence staining, n=3 for each group.  Results  Atoh1 mRNA was highly expressed in the embryonic cerebellar rhombic lip (RL) and the external granule layer (EGL). At the protein level, staining result of Atoh1-3*V5-P2A-tdTomato/+ knockin mice showed that Atoh1 was mainly expressed in the RL and the outer layer of the proliferative EGL. Atoh1-Cre;tdTomato/+   transgenic reporter mice showed that tdTomato positive cells were highly expressed in the EGL and the inner granule cell layer (IGL) during the whole granule cells   development.   Conclusion  During cerebellar development, Atoh1 mRNA and protein were highly expressed in the embryonic rhombic lip and postnatal external granule layer.    However, at the protein level, the expression of Atoh1 in the rhombic lip and external granule layer were different between knockin mice and transgenic mice.  References | Related Articles | Metrics

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Anatomy of the original short head of biceps-muscular structure XU Gao-lei LIU Jin YI Shi-qi ZHANG Yi-zheng ZHANG Zhen-hua 2023, 54 (2):  216-219.  doi: 10.16098/j.issn.0529-1356.2023.02.013 Abstract ( )   PDF (7565KB) ( )   Objective  To analyze and describe the morphological characteristics of the proximal origin of the biceps brachii, and measure the anatomical data of its tendons, so as to provide references for related clinical applications of the biceps brachii.    Methods  Six adult shoulder specimens (4 on the left and 2 on the right) were dissected to observe the origin, morphology, and course of the biceps brachii tendon and its adjacent relationship with the coracobrachialis muscle, musculocutaneous nerve and other structures. And then the length, width, and thickness of the biceps tendon were measured.    Results  The origin of the short head of biceps consisted of muscle fibers attaching directly to the tip of the coracoid process, with a thin, tendinous aponeurosis covering its anterior surface, rather than a true tendon as previously described. The mean of length，widths and thickness of the short head of biceps were (107.7±9.6) mm，(20.0±1.5)mm and (5.8±2.0)mm.    Conclusion  The short head of the biceps brachii is directly from the coracoid process with muscle fibers.  References | Related Articles | Metrics