Acta Anatomica Sinica

Exercise and complex environment inhibiting lipopolysaccharide-induced dopaminergic neuron damage in substantia nigra

CAO Cong HUANG Qin-wen WANG Hong XU Ze-ting ZHANG Chan3 SHAN Yi-wen FAN Xiao-xiao LIAO Min

2024, 55 (3): 253-259. doi: 10.16098/j.issn.0529-1356.2024.03.001

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Objective To investigate the effects of exercise and complex environment on lipopolysaccharide (LPS)-induced dopaminergic neuron death in the substantia nigra of midbrain. Methods C57BL/6 mice were divided into control group, LPS group, LPS+swimming group and LPS+complex environment group, with 7 mice in each group. The mice in the LPS group were injected with LPS into the brain to establish an inflammatory model of Parkinson ’s disease and lived in cages for 2 weeks. Mice in LPS+swimming group were forced to swim for 15 minutes every day for 2 weeks after modeling. The mice in the LPS+complex environment group were placed in a complex environment for 2 weeks after modeling. The control group mice were not treated. After 14 days of modeling, behavioral experiments such as footprint, open field and rotating rod were performed on each group of mice to detect the autonomous exercise ability, exercise balance ability and depression level of mice. The expressions of tyrosine hydroxylase (TH) in substantia nigra was detected by immunohistochemical staining and Western blotting. The expressions of brain-derived neurotrophic factor (BDNF), Caspase-3, interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) in the substantia nigra of the midbrain were detected by Western blotting. The transcription levels of IL-1β, IL-6 and TNF-α in substantia nigra were detected by RT-PCR. Results Compared with the control group, the exercise ability and balance ability of mice in LPS group, LPS+swimming group and LPS+complex environment group decreased, the depression level increased（P＜0.001）, the number of TH positive neurons and BDNF protein decreased significantly（P＜0.001）, and the contents of Caspase-3, IL-1β, IL-6 and TNF-α increased significantly（P＜0.001）. Compared with the LPS group, the exercise ability and balance ability of the mice in the LPS+swimming group and the LPS+complex environment group were restored, the depression level decreased significantly（P＜0.01）, the survival number of TH positive neurons and the content of BDNF increased significantly（P＜0.01）, Caspase-3, IL-1β, IL-6 and TNF-α reduced significantly（P＜0.01）, and the phenomenon in the LPS+complex environment group was more significant. Conclusion Exercise and complex environment can inhibit LPS-induced central nervous system inflammation in mice, thereby reducing damage to midbrain substantia nigra neurons, and the inhibitory effect of LPS+complex environment group is more significant.

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Neuromuscular electrical stimulation promoting the recovery of motor function in mice after spinal cord injury by regulating interleukin-6/signal transducer and activator of transcription-3 signaling pathway

QIN Jian-feng SONG Hai-wang SUN Bao-fei JI Yang-dan LONG Si-fang YANG Dan

2024, 55 (3): 260-267. doi: 10.16098/j.issn.0529-1356.2024.03.002

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Objective To observe the effect of neuromuscular electrical stimulation (NMES) on interleukin-6 (IL-6)/STAT3 signaling pathway in mice after spinal cord injury, and to explore the mechanism of its effect on motor function recovery. Methods Seventy-two SPF grade mice were randomly divided into sham operation group, spinal cord injury (SCI) group and NMES group. BMS score, inclined plane test and neuromuscular electrophysiology (EMG) were used to evaluate the recovery of spinal cord injury in mice. Western blotting and Real-time PCR were used to detect the expression of inflammatory factors，IL-6/STAT3, glial fibrillary acidic protein (GFAP)and brain-derived neurotrophic factor (BDNF) in spinal cord tissues of three groups of mice. HE staining was used to observe the pathological changes of spinal cord injury. Results BMS scores and the inclined plane test of mice in the NMES group were higher than those in SCI group (P<0.05). The maximum amplitude of motor evoked potential in NMES group was higher than that in SCI group (P<0.05). The expressions of TNF-α, IL-12A and GFAP in the spinal cord of NMES group were lower than that of SCI group (P<0.05), while the expressions of TGF-β, IL-10 and BDNF were higher than that of SCI group (P<0.05). The protein expressions of IL-6/STAT3 signaling pathway of NMES group were lower than that of SCI group(P<0.05). Conclusion Neuromuscular electrical stimulation plays an anti-inflammatory role by inhibiting the IL-6/STAT3 signaling pathway, thereby promoting the recovery of hind limb motor function in mice after spinal cord injury.

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Protective effect of Xuebijing on hippocampal neurons in mice with anti-N-methyl-D-aspartate receptor encephalitis and its mechanism#br#

CHEN Lin YAN Li-min XING Huai-jie CHEN Min LI Xiao-yan ZENG Chao-sheng

2024, 55 (3): 268-275. doi: 10.16098/j.issn.0529-1356.2024.03.003

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Objective To investigate the therapeutic effect of Xuebijing on anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis mice and its effect on the changes of helper T cell 1 (Th1)/helper T cell 2 (Th2). Methods The experimental groups included control group, model group, low-dose Xuebijing (low-XBJ) group, high-dose Xuebijing (high-XBJ) group, with 10 mice in each group, except the control group, the other 3 groups of mice were treated with antigen injection and immune stimulation to establish anti-NMDAR encephalitis models, the mice in the low-XBJ and high-XBJ groups were intraperitoneally injected with 5 ml/kg and 10 ml/kg of Xuebijing injection, respectively, once every 12 hours for 3 consecutive days; After 2 weeks, HE staining was used to observe the pathological changes of mice brain tissue, Nissl staining was used to observe the morphological changes of mice neurons, TUNEL staining was used to detect the apoptosis of mice neurons, ELISA was used to detect the contents of tumor necrosis factor-α (TNF-α), interleukin(IL)-1β, IL-4 and interferon-γ (IFN-γ) in serum and cerebrospinal fluid of mice, immunohistochemical staining and Western blotting were used to detect the expression of IFN-γ and IL-4 in brain tissue, flow cytometry was used to detect the distribution of Th1 and Th2 cells in peripheral blood, and the ratio of Th1/Th2 was calculated. Results Compared with the model group, hippocampal tissue damage was improved in low-XBJ group and high-XBJ group, the morphological changes of neurons were small, the morphology of Nissl bodies was more complete and the number of Nissl bodies was increased, the TUNEL positive rate of neurons decreased, the contents of TNF-α, IL-1β and IFN-γ in serum and cerebrospinal fluid decreased while the content of IL-4 increased, the percentage of IFN-γ positive cells and the relative expression of protein in brain tissue decreased, the percentage of IL-4 positive cells and the relative expression of protein increased, the proportion of cluster of differentiation (CD)4⁺IFN-γ⁺ labeled Th1 cells and the ratio of Th1/Th2 in peripheral blood decreased, and the proportion of CD4⁺IL-4⁺ labeled Th2 cells in peripheral blood of mice in high-XBJ group increased, the differences were statistically significant (P<0.05). In addition, compared with the low-XBJ group, the improvement effect of the above detection indicators in the high-XBJ group was more obvious, and the differences were statistically significant (P<0.05). Conclusion Xuebijing can improve hippocampal neuronal damage in anti-NMDAR encephalitis mice, which may play a therapeutic role by reducing the expression of IFN-γ, promoting the expression of IL-4 and maintaining the balance of Th1/Th2 cells.

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Heshouwuyin delaying the aging of Leydig cells in rat testis through DNA methyltransferase 1

WU Tian LU Shuo-ya YANG Yu-jiao DUAN Yu-lei AN Qi-chao ZHEN Xiao-lan NIU Si-yun

2024, 55 (3): 276-284. doi: 10.16098/j.issn.0529-1356.2024.03.004

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Objective To investigate whether Heshouwuyin can delay the aging of Leydig cells in rat testis through DNA methyltransferase 1 (DNMT1). Methods Totally 40 male Wistar rats were randomly divided into 4 groups, with 10 rats in each group. Immunohistochemistry was used to detect the expression levels of DNMT1 in testis tissue of rats. Testosterone content in serum of rats in each group was detected by ELISA test. A rat Leydig cell aging model was established by free radical oxidative damage. DNMT1 was knocked down by lentivirus in Leydig cells, and the cell senescence status and the testosterone content and testosterone synthesis key enzyme 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 family member 11A1 (CYP11A1) content secreted by cells were detected by β-GALactosidase（β-GAL）staining, immunofluorescenct staining and ELISA. Results Compared with the young control group (YCG), the expression of P16 protein and the positive rate of β-GAL in the testis tissue of rats in the natural aging group (NAG) increased significantly, and the expression of DNMT1 and serum testosterone levels decreased(P<0.05). However, after Heshouwuyin intervention, the expression of P16 protein and the positive rate of β-GAL in the testis of aging rats were reduced, and DNMT1 expression and the serum testosterone levels increased (P<0.05). The same trend was observed in Leydig cells. Knockdown of DNMT1 in Leydig cells, β-GAL positivity and P16 protein expression increased significantly, and testosterone secretion and testosterone synthesis key enzymes 3β-HSD, CYP11A1 content from Leydig cells decreased significantly, compared with the normal control group (NCG) (P<0.05). When Heshouwuyin was added, the above phenomenon was improved. Conclusion Heshouwuyin can delay the aging of rat Leydig cells through DNMT1.

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College of Veterinary Medicine, Shanxi Agricultural University, Shanxi Taigu 030801, China

LU Jia-yin YAO Zhi-chao HAO Xiao-jing YAN Yi MA Pei ZHANG Hui-ling WANG Hai-dong

2024, 55 (3): 285-294. doi: 10.16098/j.issn.0529-1356.2024.03.005

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Objective To investigate the dynamic expression with the time change of N6-methyladenosine（m6A） methylation-related factors in the repair process of skeletal muscle injury and its mechanism in the inflammatory response of macrophage in the injure process. MethodsIn vivo mice models of BaCl2 injury in the gastrocnemius were established. Four mice per group in the control group and injury group. Gastrocnemius tissues were harvested at day 1, 3, 5, 7, and 9 after injury for experiments. Primary gastrocnemius muscle tissue cells, muscle satellite cells, muscle cells, and cell line C2C12 cells were treated with dexamethasone (DEX, 50 μmol/L) to mimic injury. Lipopolysaccharide (LPS, 100 μg/L) induced RAW264.7 cell lines to mimic the inflammatory response after skeletal muscle injury, and STM2457 (30 μmol/L) was added to inhibit the effect of methyltransferase 3 (Mettl3) before LPS treatment. The expression of m6A methylation-related factors (Writers, Erasers, Readers) and inflammation factors were detected by Real-time PCR and Western blotting. Results The muscle fibers were dissolved and then gradually repaired with the extension of injury time, the number of monocytes/macrophages increased first and then decreased, and the Pax7 mRNA level increased first and then decreased with the change of injury time. Compared with the control group, the mRNA and protein levels of m6A methylationrelated factors in gastrocnemius did not change significantly on the injury-1 day. However, they were significantly increased on the injury-3 days compared with the control group (P<0.05), and then obviously decreased on the injury-5 days group compared with the injury-3 days group(P<0.05). Compared with the control group, they were no significant differences on the injury-7 days group and-9 days group.In vitro DEX decreased the mRNA levels of m6A methyltransferase factors in primary muscle satellite cells and C2C12 cells and increased the mRNA expression level of methylation-recognition enzyme factors (P<0.05). The mRNA levels of m6A methylation-related factors increased significantly in skeletal muscle tissue cells and myocytes after DEX treatment (P<0.05). After LPS treatment, the mRNA and protein expression levels of m6A methylation-related factors and the mRNA expression levels of inflammatory factors interleukin(IL)-6 and IL-1β in macrophages increased significantly (P<0.05), while the levels of IL-6 and IL-1β mRNA in macrophages decreased significantly when the Mettl3 was inhibited (P<0.05). Conclusion m6A methylation-related factors primarily is activated in the damaged muscle cells and inflammation response of macrophages. Inhibition of m6A methyltransferase can reduce the inflammatory response of macrophages.

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Roles of hepatic Dishevelled/Egl-10/pleckstrin domain-containing protein 5/mammalian target of rapamycin complex 1 signaling axis on the development of non-alcoholic fatty liver disease

XU Lin XIONG Xi-wen LI Zun HUANG Rong MA Hong-hui MA Jie

2024, 55 (3): 295-301. doi: 10.16098/j.issn.0529-1356.2024.03.006

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Objective To investigate the effect of hepatic Dishevelled/Egl-10/pleckstrin domain-containing protein 5(DEPDC5)/mammalian target of rapamycin complex 1(mTORC1) on nonalcoholic fatty liver disease by establishing a high-fat diet feeding model of Depdc5 gene hepatocyte specific knockout mice. Methods Depdc5flox/flox mice were constructed and mated with Alumin-Cre mice to obtain Depdc5flox/flox;Alb-Cre mice (LKO), Depdc5flox/flox mice were as control (Loxp). Totally 32 male mice aged 2-3 months were randomly divided into high-fat-diet LKO group, high-fat-diet Loxp control group, high-fat-diet + rapamycin LKO group, and high-fat-diet + rapamycin Loxp control group, with 8 mice in each group. Liver serum biochemistry, lipid content, protein, mRNA and pathological sections were detected; Graphpad prism 8 software was used for statistical analysis. Results High-fat-diet induced liver steatosis in Loxp mice, while LKO mice were protected from steatosis but had aggravated liver injury. Rapamycin treatment attenuated the hyperactivation of mTORC1 pathway caused by Depdc5 knockout, alleviated the liver steatosis in Loxp mice and liver injury in LKO mice. Conclusion Deletion of Depdc5 gene protects mice from high-fat-diet induced liver steatosis and rapamycin treatment might be used to improve liver injury caused by DEPDC5 loss of function.

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Osteoblast-derived exosome mediating the effect of microRNA-494 on bone metabolism and bone remodeling balance in osteoporotic rats

LIN Wei LI Chao-yi TANG Jie ZHANG Pi-jun

2024, 55 (3): 302-310. doi: 10.16098/j.issn.0529-1356.2024.03.007

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Objective To investigate the effect of osteoblastderived exosome (Exo) mediating microRNA (miR)-494 on bone metabolism and bone remodeling balance in osteoporosis (OP) rats and its mechanism. Methods Exosomes was isolated and identified from MC3T3-E1 osteoblast cell line and transferred to Exo by electrical transfer. Forty SD rats were randomly divided into control, model, miR-494 mimic (miR-494), and miR-494 inhibitor group, 10 rats in each group. The ovaries were removed to construct the OP model except the control group. After modeling, the miR-494 group and miR-494 inhibitor group received tail vein injections of exosomes containing the corresponding miRNA, at a dose of 3×109 particles. Four weeks later, bone parameters were detected in each group of rats by Micro-CT, serum bone markers were measured by ELISA, pathological changes in bone tissue were observed by HE staining, osteoclast numbers were detected by tartrate-resistant acid phosphatase (TRACP) staining, and the expression levels of bone remodeling-related proteins and toll-like receptor 4(TLR4) pathway-related proteins were determined by Western blotting. Results Typical cup-shaped or round exosomes were successfully isolated with a diameter of about 100 nm from MC3T3-E1 cells, which contained CD63, CD9, tumor susceptibility gene 101(TSG101), heat shock protein 70(HSP70) proteins and can be taken up by MC3T3-E1 cells. Compared with the model group, the bone parameters of the rats in the miR-494 mimic group decreased, serum bone markers bone Gla protein (BGP), TRACP, C-terminal telopeptide of type Ⅰ collagen (CTX-Ⅰ) increased, osteoprotegerin (OPG), procollagen type Ⅰ N-terminal propeptide (PⅠNP) decreased, bone trabeculae structure was disordered, osteoclasts increased, bone morphogenetic protein 2(BMP-2), Runt related transcription factor 2(RUNX2) in bone tissue downregulated, receptor activator of nuclear factor kappa-B ligand (RANKL) upregulated, TLR4, nuclear factor kappa-B p65(NF-κB p65) and myeloid differentiation primary response 88(MyD88) upregulated (all P<0.05). In contrast, the situation of the miR-494 inhibitor group was opposite, bone parameters and OPG, PⅠNP increased, BGP, TRACP, CTX-Ⅰ decreased, bone structure returned to normal, osteoclasts decreased, BMP-2, RUNX2 in bone tissue upregulated, RANKL downregulated, TLR4, NF-κB p65 and MyD88 downregulated (all P<0.05). Conclusion The transfer of miRNA-494 by Exo aggravates abnormal bone metabolism in OP rats and inhibits bone remodeling balance, suggesting that the mechanism of action may be related to the regulation of TLR4 pathway.

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Screen of key characteristic genes and analysis of immune cell infiltration in metastatic nasopharyngeal carcinoma base on machine learning#br#

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LU JinCHEN Yun-fanZHANG Hao-xuanHUANG Xue-ying

2024, 55 (3): 311-318. doi: 10.16098/j.issn.0529-1356.2024.03.008

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Objective To screen the key characteristic genes of metastatic nasopharyngeal carcinoma (mNPC) and analyze the immune cell infiltration in tumor microenvironment using machine learning algorithm. Methods Firstly, the training set GSE103611 was downloaded from the GEO database, and the data were subjected to differential expression gene (DEGs) screening, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genome (KEGG) and immune cell infiltration analysis. Second, the predicted genes in DEGs were screened by least absolute shrinkage and selector operation( LASSO) regression, and the characteristic genes were screened by using the expression level of the predicted genes and receiver operating characteristic(ROC). Third, the correlation between characteristic genes and immune cells was further analyzed to determine the key characteristic genes. Finally, the expression levels of key characteristic genes and ROC were verified using the reverse validation set GSE1245 data. Results A total of 136 DEGs were obtained, and their KEGG were mainly enriched in cytochrome P450, tumor necrosis factor(TNF) signaling pathway, prion disease, EB virus infection, and other pathways. GO was mainly enriched in the negative regulatory processes of peptide-based tyrosine phosphorylation modification, viral gene expression, and B cell and leukocyte activation. The difference in the degree of infiltration of the 22 immune cells in nasopharyngeal carcinoma(NPC) and mNPC was not significant. Two key characteristic genes (DAZ1 and SCCPDH) of mNPC were finally obtained by LASSO regression, and they were significantly correlated with immune cells in the mNPC microenvironment (P<0.05). In the reverse validation data set, the differential expressions of DAZ1 and SCCPDH between non\|NPC(nNPC) and NPC groups were not significant (P>0.05), and the AUC values of ROC of both were less than 0.6. Conclusion DAZ1 and SCCPDH are the key characteristic genes of mNPC and can be used as important markers for mNPC and immunotherapy.

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Construction and validation of a prognostic model of hepatocellular carcinoma based on immune-related genes

CHEN Don-dong LOU Jin-jin HUANG Yan-yan ZHOU Lu LI Shi-bo XU Li-yun

2024, 55 (3): 319-328. doi: 10.16098/j.issn.0529-1356.2024.03.009

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Objective To construct a prognostic model for liver hepatocellular carcinoma(LIHC) based on immune-related genes. Methods LIHC and normal tissue samples were downloaded from the UCSC Xena database and The Cancer Genome Atlas (TCGA) database. Differential analysis was performed on the gene data of LIHC samples and adjacent/normal samples. Enrichment analysis was conducted on differentially expressed genes. Kaplan-Meier survival analysis was performed on liver cancer samples from the TCGA cohort to obtain survival- and immune-related differentially expressed genes. LASSO Cox and multivariate Cox regression analysis were used to identify hub genes and construct a gene risk prognostic model. Data from a high-throughput gene expression (GEO) database was obtained for external validation. The sensitivity of hub genes to common anticancer drugs was investigated using the CellMiner database. ResultsEnrichment analysis result indicated that differentially expressed genes were mainly associated with metabolic pathways. Through differential analysis and Kaplan-Meier survival analysis, 25 survival- and immune-related differentially expressed genes were obtained. Based on the result of LASSO Cox and multivariate Cox regression analysis, five hub genes (FYN, CSF3R, HLA-G, FOS, BIRC5) were identified and a nomogram was constructed. The concordance index(C-index) value for the training cohort and validation cohort were 0.739 and 0.625, respectively. Based on the sensitivity of hub genes to anticancer drugs, 12 types of anticancer drugs were selected for subsequent experiments. Conclusion This model can effectively predict the prognosis of LIHC patients and provide a new insights for immune therapy in LIHC.

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Observation and imaging analysis of signs of ankylosing spondylitis in spinal specimens#br#

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ZHONG Wei-xing WANG Zhi-hong LI Jun-hua LIAO Li-qing CHEN Zu-jiang LI Yi-kai

2024, 55 (3): 329-333. doi: 10.16098/j.issn.0529-1356.2024.03.010

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Objective To provide anatomical, radiological, and clinical diagnostic and therapeutic references for ankylosing spondylitis and spinal surgical operations. Methods Non-measurement spinal observations, X-ray examinations, and measurements were performed on two spinal specimens, along with digital image acquisition and processing. Results The first specimen included thoracic vertebra 7 (T7) to lumbar vertebra 3 (L3), with an average total length of 29.7 cm; the second specimen ranged from cervical vertebra 7 (C7) to lumbar vertebra 2 (L2), with an average total length of 38.3 cm. The specimens showed partial or complete calcification of ligaments, ossification of the small joints and intervertebral discs, and osteoporosis; The anterior-posterior diameter (width) of the vertebral foramen was narrower than that of a normal adult, while most of the superior-inferior diameter (height) was wider. Radiographically, the anterior longitudinal ligament calcification appeared as dot-like or striated, but it was actually flaky in the actual specimens. The specimens provided views of the facet joints, costovertebral joints, and intervertebral foramina that was difficult to demonstrate on two-dimensional X-ray images. Conclusion As ankylosing spondylitis progresses, the range of motion in spinal bending and rotation decreases, as does the extent of thoracic expansion, thereby affecting respiration and complicating procedures such as intraspinal anesthesia and sacral canal injections. In terms of diagnosis, bone specimens and X-ray films allow us to understand the development process and severity of ankylosing spondylitis more directly and accurately.

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Digital optimization design combined with 3D printing technology for bone tunnel creation in distal tibiofibular syndesmosis injury

LI Ming-hui XIONG Ji-xiang ZHOU Xin ZHANG Lei

2024, 55 (3): 334-338. doi: 10.16098/j.issn.0529-1356.2024.03.011

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Objective To employ digital optimization design in the quest for the optimal placement position of the distal tibiofibular syndesmosis tunnel. Methods A total of 100 distal tibiofibular syndesmosis CT datasets, acquired from 95 patients with syndesmosis injuries the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, were imported into the Mimics 19.0 software for the purpose of reconstructing models. Created virtual tunnels between the tibia and fibula and overlaid them on the reconstructed models as virtual models. Additionally, 3D printing was used to create actual models. In this study, eight parameters were measured. These included the length of the bone tunnel, the distance from the center of the tibial tunnel to the anterior and posterior aspects of the tibia, the distance from the center of the fibular tunnel to the anterior and posterior aspects of the fibula, the distance from the tibial tunnel point to the medial malleolus, the distance from the fibular tunnel point to the lateral malleolus, and the angle between the bone tunnel and the horizontal plane. Results Eight parameters were compared between the virtual model and the actual model, and there was no significant difference in the all measurement results of the actual bone tunnel and the virtual bone tunnel (P>0.05). The anatomical analysis revealed a statistically significant difference in gender (P<0.05), while there was no significant statistical difference observed in the affected side (P>0.05). Conclusion Digital optimization design can provide precise and reliable bone tunnels for elastic fixation of the tibiofibular syndesmosis injury, thereby providing good stability for the surgery and helping patients recover ankle joint function as soon as possible.

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Finite element analysis of titanium rods after vertebral column decancellation osteotomy for ankylosing spondylitis

SU Bao-ke WANG Yong-qing KANG Zhi-jie WANG Hai-yan JIN Feng LI Xiao-he CAO Zhen-hua HUO Jia-xuan ZHU Yong LI Feng

2024, 55 (3): 339-344. doi: 10.16098/j.issn.0529-1356.2024.03.012

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Objective To analyze the stress changes of thoracic vertebra (T)11-sacrum (S) titanium rods in patients with ankylosing spondylitis after vertebral column decancellation (VCD) osteotomy, and provide reference for the selection and improvement of titanium rods before surgery. Methods The original data of the continuous scanning tomographic images of patients with ankylosing spondylitis after VCD osteotomy were imported into Mimics 21.0 in DICOM format, and T₁₁-S vertebrae, screws and titanium rods were respectively reconstructed. They were imported into 3-Matic to establish a preliminary geometric modeling, and then processed with noise removal, paving, smoothing, etc. The improved model was imported into Hypermesh 10 software for grid division, and the material was imported into ANSYS 19.2 to display the finite element model after attribute assignment, Set the boundary and load conditions, and measure the stress value at the connection between the screw and the titanium rod. Results Under neutral position, forward bending, lateral bending, and axial rotation conditions, the titanium rod had the highest stress at the upper vertebrae (T₁₁) and the lowest stress at the top vertebrae (L₃); Under the backward extension condition, the titanium rod has the highest stress at the lower end vertebra (L₅). Conclusion In the upper and lower vertebrae, it is possible to consider increasing the diameter of the titanium rod, enhancing its hardness, or changing it to a double rod.

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Effect of sperm DNA fragmentation on in vitro fertilization-embryo transfer outcome among middle-aged men

WANG Ping WANG Jing CHEN Xian-bing CHEN Xi

2024, 55 (3): 345-348. doi: 10.16098/j.issn.0529-1356.2024.03.013

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Objective To explore the relationship between sperm DNA fragmentation index(DFI) and sperm quality, as well as in vitro fertilization-embryo transfer (IVF-ET) preganancy outcomes in middle-aged men. Methods A total of 180 semen specimens from males aged over 38 years who received comventional IVF-ET treatment were collected and divided into two groups(<30% and ≥30%) based on the DIF threshold. Main measures result including: normal semen parameters, hormone levels, DFI, fertilization rate, good quality embryo rate and pregnancy rate. Results DFI was closely related to male follicle-stimulating hormone（FSH）, sperm motility，morphology, and the sperm vitality decreased with increasing DFI level (P<0.05). When DFI≥30%, the rate of high-quality embryos decreased (P<0.05), but there was no significant difference in the pregnancy rate (P>0.05). Conclusion DFI can serve as an important indicator for routine analysis of semen in middle-aged men. DFI affects the rate of high-quality embryos, but it is not related to the clinical pregnancy outcome of assistant reproduction technology (ART).

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Analysis of human composition of the Gannan Tibetans

SUN Si-yu LI Yong-lan ZHENG Lian-bin YU Ke-li

2024, 55 (3): 349-355. doi: 10.16098/j.issn.0529-1356.2024.03.014

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Objective To reveal the characteristics of Tibetan human body composition in Gannan. Methods The body composition of 410 Tibetan adults (221 males and 189 females) in Gannan was measured by bioelectrical impedance method. The data were processed by SPSS 20.0 statistical software. Results Related analysis results showed that the total body muscle mass, estimated fat level and trunk muscle mass of the male decreased with age, while the percent body water, visceral fat level and percent trunk fat increased with age. The stature and left lower limb muscle mass of the female decreased with age, while body weight, percent body fat, body mass index, visceral fat level, percent right upper limb fat, right upper limb muscle mass, percent left upper limb, left upper limb muscle mass, percent right lower limb fat, percent left lower limb fat, left lower limb muscle mass and percent trunk fat increased with age. Analysis of single factor variance showed that there were statistically significant differences in the four indexes of male percent body water, visceral fat level, percent trunk fat and trunk muscle mass, while there were no statistically significant differences in the other 14 indexes among age groups. Females weight, stature, percent body fat, total body muscle mass, estimated fat level, body mass index, visceral fat level, percent right upper limb fat, right upper limb muscle mass, percent left upper limb fat, left upper limb muscle mass, percent right lower limb fat, percent left lower limb fat, left lower limb muscle mass, percent trunk fat, trunk muscle mass 16 index values were statistically significant differences between age groups, the remaining two index values were not statistically significant differences between age groups. The results of u test showed that the body weight, total body muscle mass, estimated fat level, percent body water, limb muscle mass and trunk muscle mass of males were higher than those of females, and percent body fat, limb fat rate, visceral fat rate and trunk fat rate of females were higher than those of males. According to the body mass index, Gannan Tibetan male and female were overweight; according to the percent body fat, Gannan Tibetan males were nonobesity and females were obesity. Conclusion The body mass index and overall fat content of Tibetan males and females in Gannan are relatively high among the 13 ethnic groups.

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Body composition and obesity of Ewenki, Daur and Mongolian Buryat #br#

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XI Lu-ge YU Hui-xin LIAN Yi YI La-na HAN Yuan LI Yong-lan

2024, 55 (3): 356-362. doi: 10.16098/j.issn.0529-1356.2024.03.015

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Objective To analyze the characteristics of adult body composition and obesity status of three ethnic groups: Ewenki, Daur and Mongolian Buryat. Methods The bioelectrical impedance analysis（BIA）was used to measure 18 body composition components in three adults ethnic groups: 245 (male 124, female 121) Ewenki, 207 (male 90, female 117) Daur, and 181 (male 74, female 107) Mongolian Buryat . The data were processed using Excel 2016 and SPSS 24.0 statistical software. Results The result of correlation analysis showed that visceral fat level was significantly and positively correlated with age (P＜0.01). Stature, total body muscle mass, estimated bone mass and trunk muscle mass were all significantly and negatively correlated with age (P＜0.01) in males and females of the three ethnic groups. The percent body fat, percent left upper limb fat and percent trunk fat were positively correlated with age in Ewenki males (P<0.05 or P＜0.01) and the percent body fat, body mass index (BMI), percent limb fat and percent trunk fat were positively correlated with age in Ewenki females (P<0.05 orP>0.01). Body weight, BMI, percent left upper limb fat, left upper limb muscle mass, bilateral lower limb fat and muscle mass were all negatively correlated with age in Daur males (P<0.05 or P>0.01 ). Body weight, upper limb muscle mass and left lower limb muscle mass were negatively correlated with age (P<0.05 or P＜0.01) in Mongolian Buryat males and percent trunk fat was positively correlated with age (P<0.05) in Mongolian Buryat females. Comparison between ethnic groups showed that most of the body composition index values of the Ewenki and Mongolian Buryat divisions were closest to each other, and the body fat content was higher than that of the Daur. BMI, percent body fat and visceral fat level were all manifested in the Mongolian Buryat Department>Ewenki>Daur. Cluster analysis showed that Ewenki, Daur and Mongolian Buryat were closer to the northern groups and further from the southern groups. Conclusion The Ewenki is most similar to the Mongolian Buryat in body composition characteristics all three ethnic groups has a more serious obesity problem.

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Optimisation of CUBIC tissue clearing technology based on perfusion methods

GONG Chuan-hui QIU Jia-yi YIN Ke-xin ZHANG Ji-ru HE Cheng YUAN Ye LÜ Guang-ming

2024, 55 (3): 363-370. doi: 10.16098/j.issn.0529-1356.2024.03.016

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Objective In order to shorten the transparency time of clear, unobstructed brain imaging cocktails and computational analysis(CUBIC), improve the transparency efficiency, and explore the possibility of applying hydrophilic tissue transparency technique, this study was conducted to optimize the perfusion of CUBIC technique and compare it with four hydrophilic tissue clearing method in terms of tissue transparency effect, transparency time, area change, volume change and adeno-associated virus (AAV) fluorescence retention. Methods Brain, liver, spleen and kidney of 6 adult Institute of Cancer Research(ICR) mice were subjected to clearing treatment by SeeDB, FRUIT, ScaleS and CUBIC method, respectively. The area and gray value of the samples were measured by Image J 1.8.0, and the volume before and after transparency was measured by drainage method to compare the transparency effect, time and size deformation of each group. Perfusion optimization of the CUBIC was performed by improving the perfusion rate with the optimal perfusion dose, each group of the experimental sample size was 6. Fluorescence preservation by different techniques was evaluated by injecting AAV in the motor cortex of 16 adult mice and taking the cervical spinal segments for transparency treatment after four weeks, and the fluorescence photographs were measured by Image J 1.8.0 to measure the mean fluorescent intensity. Results The optimal perfusion rate and dose of CUBIC was 15 ml/min and 200 ml respectively. For transparency ability and speed, the perfusion CUBIC had the lowest mean gray value and took the shortest time, while CUBIC consumed the longest time, and SeeDB, FRUIT, and ScaleS did not show good transparency ability. In terms of area and volume changes, several techniques showed different degrees of expansion after transparency of tissues or organs. In terms of fluorescence retention, perfusion CUBIC showed the best retention of green fluorescent protein (GFP) fluorescence signal, followed by CUBIC, ScaleS, FRUIT, and SeeDB. Conclusion Perfusion CUBIC technique shows the best tissue transparency, the shortest transparency time, and the most AAV fluorescence retention compared with other techniques.

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Research progress on myelinassociated inhibitors and their receptors in spinal cord injury repair

LIU Xin-tong PENG Qing-hua LIU Huai-cun ZHANG Wei-guang

2024, 55 (3): 371-377. doi: 10.16098/j.issn.0529-1356.2024.03.017

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Spinal cord injury is a traumatic disease, commonly seen in falling injuries, traffic accidents, heavy injuries, etc, which could cause motor, sensory and autonomic dysfunction below the level of spinal cord injury. Myelin-associated inhibitors play a role in promoting the collapse of growth cones and inhibiting axonal regeneration in the injured spinal cord microenvironment, which is the main reason for the difficult repair of spinal cord injury. Myelinassociated inhibitors (MAIs), such as neurite outgrowth inhibitor (Nogo), oligodendrocyte-myelin glycoprotein (OMgp) and myelin-associated glycoprotein (MAG), along with their receptor proteins, such as Nogo-A/Nogo-66 receptor 1 (NgR1), paired immunoglobulin-like receptor B (PirB), sphingosine-1-phosphate receptor 2 (S1PR2), are the important regulatory factors in the spinal cord microenvironment. They can inhibit therepair process of spinal cord injury by affecting the signaling pathway of neuron axon growth. Although the mechanism of spinal cord injury repair is still unclear, the regulation of myelin-related inhibitory factor proteins and downstream signaling pathways remain an important therapeutic approach for spinal cord injury. In this paper, the role of MAI proteins and their receptors in spinal cord injury repair in recent years were reviewed to provide a new target for spinal cord injury repair and provide more ideas for clinical treatment after spinal cord injury.

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