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    Neurobiology
    Proteomic analysis of hippocampal tissue in rats with chronic diffuse axonal injury based on 4D-Label-free technique
    ZHANG Li XIONG Hong-li ZHU Shi-sheng
    2024, 55 (5):  515-523.  doi: 0.16098/j.issn.0529-1356.2024.05.001
    Abstract ( )   PDF (3802KB) ( )  
    Objective   To screen differentially expressed proteins (DEPs) in the hippocampal tissues of rats with chronic diffuse axonal injury (DAI), in order to provide an important theoretical basis for exploring the potential pathogenesis of chronic DAI as well as for clinical diagnosis, screening of drug therapeutic targets, and assessment of prognosis.  Methods   The experimental animals were divided into model group (DAI group, n=20) and control group (CON group, n=20). A modified Marmarou method  was used to establish a DAI model in SD rats. After three weeks of modeling, the changes in protein profiles of hippocampal tissues between two groups were compared with the 4D-Label-free technology, and DEPs were screened by the fold change(FC) in expression of the DAI group/CON group of >1.2 or <0.83 with P<0.05. Bioinformatical analysis of selected DEPs was performed by using gene ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genes (KEGG) pathway enrichment analysis method.  Results   A total of 92 DEPs were screened with 52 upregulated and 40 downregulated. The result  of GO analysis showed that DEPs were mainly involved in biological process (BP) functions such as dephosphorylation, ATP synthesiscoupled electron transfer, positive regulation of hydrogen peroxide-mediated programmed cell death, and internalization of neurotransmitter receptors. The results of KEGG pathway analysis suggested that DEPs were mainly involved in signaling pathways such as metabolic pathways, reactive oxygen species, neurodegenerative pathwaysmultiple diseases, intraretrograde cannabinoid signaling, and glutathione metabolism.  Conclusion   The DEPs in hippocampus of chronic DAI group rats are screened by 4D-Label-free technique. The selected DEPs and their enriched BP and signaling might pathway provide theoretical basis for further study of chronic DAI. 
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    Improvement of depression-like behaviors by treadmill exercise combined with black lycium polysaccharide in mice
    CHEN Wei CHEN Jia-qin WANG Yi-rong
    2024, 55 (5):  524-532.  doi: 10.16098/j.issn.0529-1356.2024.05.002
    Abstract ( )   PDF (6185KB) ( )  
    Objective   To study the effects of 7week treadmill exercise combined with black lycium polysaccharide on depression-like behaviors and explore the alterations of hippocampal α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor (AMPAR)associated pathways in mice.  Methods   Fifty male Kunming (KM) mice were randomly divided into blank group (K, n=10) and chronic unpredictable mild stress group(CUMS, n=40). For the CUMS group, 13 random chronic unpredictable mild stress method  were used to induce depression-like behaviors. After successful modeling, the mice were further divided into model group (M), treadmill exercise group (E), black lycium polysaccharide group (L), and treadmill exercise combined with black lycium polysaccharide group (EL); each group contained 10 mice, and intervention lasted for 7 weeks. The behavioral assessment was performed; the contents of 5-hydroxytryptamine(5-HT), brain-derived neurotrophic factor (BDNF) and dopamine (DA) in hippocampal tissue, and the levels of S100 calcium binding protein B (S100B) and neuron-specific enolase (NSE)in serum were detected; Nissl staining was carried out to observe the structure of brain tissue; AMPAR, glutamate receptor 1(GluR1) and calcium/calmodulin-dependent protein kinase α (CaMKⅡα) proteins in hippocampal tissue were detected; Real-time PCR was used to verify the expression of hippocampal AMPAR, GluR1 and CaMKⅡα mRNA obtained by mRNA sequencing.  Results   1. Compared with the blank group, the mice in CUMS group had obvious depression-like behaviors (P<0.01); the contents of 5-HT, BDNF and DA in hippocampal tissue decreased significantly, while those of S100B and NSE increased (P<0.01); Neurons in the prefrontal cortex were absent, the Nissl nacleus was condensed, and the arrangement was irregular and sparse. 2. Compared with the model group, the behavioral assessment of the mice in the E, L and EL groups were significantly improved, reflecting that the contents of 5-HT, BDNF and DA in hippocampal tissue increased, and the serous S100B and NSE levels decreased (P<0.01 or P<0.05); for sucrose preference test (SPT) and forced swimming test (FST), S100B and NSE content result  showed that treadmill exercise and black lycium polysaccharide intervention had a synergistic effect on behavioral improvement. 3. Compared with the model group, the contents of AMPAR, GluR1, and CaMKⅡα proteins in hippocampal tissue of mice in the E, L and EL groups increased (P<0.01), and there was a synergistic effect between treadmill exercise and black lycium polysaccharides; the Illumina high-throughput sequencing data showed that there were 49 upregulated genes and 18 down-regulated genes involving learning and memory, and neuronal damage repair, etc. which were closely related to postsynaptic membrane location of AMPA receptors and synaptic long-term potentiation and long-term depression, The increased transcriptions of AMPAR, GluR1 and CaMKⅡα revealed by the sequencing were verified by Real-time PCR data, and there was a synergistic effect between the treatments of treadmill exercise and black lycium polysaccharides.  Conclusion   Seven weeks of treadmill exercise and administration of black lycium polysaccharides can improve depression-like behaviors in mice, possibly by affecting the phosphorylation of CaMKⅡα to regulate AMPAR activity, promote neuronal damage repair and improve synaptic plasticity and function.
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    Exercise intervention alleviating learning and memory dysfunction of Alzheimer’s disease model mice through modulating autophagy of hippocampal neurons#br#
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    MU Lian-wei HAN Peng YANG Yun-jie
    2024, 55 (5):  533-540.  doi: I0.16098/j.issn.0529-1356.2024.05.003
    Abstract ( )   PDF (3642KB) ( )  
    Objective   To investigate the effects of exercise on autophagy of the hippocampal neurons and learning and memory abilities in APP/PS1 transgenic Alzheimer’s disease(AD)model mice.  Methods   Four-month-old male APP/PS1 mice were randomly divided into AD model sedentary group (AS) and AD model exercise group (AE). C57BL/6 mice of the same age were used as control group (CS), with 12 mice in each group. Among them, AE group mice underwent regular treadmill exercise at 10 m per minute(the first 10 minutes) and 12 m per minute(the following 50 minutes) for 60 min, 0° slope, 5 days/week, for 8 weeks. Morris water maze, novel object recognition test, immunohistochemistry, Western blotting, transmission electron microscopy, and Nissl staining were used to detect learning memory abilities, amyloid β-protein (Aβ), human microtubule-associated protein light chain 3B (LC3B), Beclin1, p62 protein expression, autophagy lysosomes and neuron number.  Results   Eight weeks of exercise intervention could significantly improve the learning and memory abilities of AD model mice in the Morris water maze and novel object recognition test. The expression levels of Aβ plaques and p62 protein in the hippocampus of mice in the AS group increased significantly, while the levels of LC3B, Beclin1, autophagy lysosomes and neurons decreased significantly. Exercise intervention significantly reduced the expression levels of Aβ plaques and p62 protein in the hippocampus of AD model mice, and increased the levels of LC3B, Beclin1, autophagolysosomes and neurons.  Conclusion   Exercise could effectively improve the damage of neuronal autophagy in the hippocampus of AD model mice and protect the neurons through promoting autophagy lysosomes to degrade pathological Aβ in the hippocampus. These result might underlie the mechanisms for the treadmill exercise-induced improvement of learning and memory abilities. 
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    Neurobiology
    Effects of microplastics exposure on learning and memory in mice and its mechanism
    JIANG Xin-ze LIU Qiang SUN Xu HOU Jiang-shan MA Rui CHENG Mei WU Yulong
    2024, 55 (5):  541-546.  doi: 10.16098/j.issn.0529-1356.2024.05.004
    Abstract ( )   PDF (3391KB) ( )  
    Objective   To investigate the effect of microplastic (MPs) exposure on learning and memory in mice, and its mechanism by observing the protein expression of brain-derived neurotrophic factor (BDNF) /tyrosine receptor kinase B (TrkB) /N-methyl-D-aspartate receptor subtype 2B (NR2B) signaling pathway and neurogenesis.  Methods  Forty male C57BL/6 mice were randomly divided into control group (Ctrl) and microplastics exposure group (MPs). Mice in MPs group were treated with 0.3 mg/(kg·d) microplastics, administered by gavage at a volume of 200 μl for 30 consecutive days. Morris water maze test was used to evaluate the spatial learning and memory ability of mice. Western blotting was used to detect the protein levels of BDNF, TrkB and NR2B in hippocampus of mice. Immunofluorescent staining was used to observe the number of doublecortin (DCX) and neuronal nuclei antigen (NeuN) positive cells in the hippocampus of mice to evaluate hippocampal neurogenesis.  Results  Compared with the control group, the ability of learning and memory decreased significantly in MPs group mice (P<0.01). The expression levels of BDNF, TrkB and NR2B in the hippocampus of MPs group mice were significantly lower than those of control group (P<0.05). The number of DCX and NeuN positive cells in the hippocampus of MPs group was significantly lower than that of control group (P<0.01).  Conclusion   MPs exposure induces learning and memory impairment which may be related to inhibiting BDNF/TrkB/NR2B signaling pathway and reducing hippocampal neurogenesis. 
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    Electroacupuncture inhibiting LPS-induced chronic neuroinflammation by regulating the cortical NF-κB/NOD-like receptor protein 3 signaling pathway
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    WANG Li-juan GAO Ce ZHAO Zhi-hong HAI Zhen LI Wen-hui HAN Qiu-qin
    2024, 55 (5):  547-555.  doi: 10.16098/j.issn0529-1356.2024.05.005
    Abstract ( )   PDF (5108KB) ( )  
    Objective   To observe the effect of electric stimulation on nuclear factor-κB (NF-κB)/ NOD-like receptor protein 3 (NLRP3) signaling pathway and microglial cell morphology in mice with lipopolysaccharide(LPS)induced chronic neuroinflammation, and to explore the protective mechanism of electric stimulation on brain of mice.  Methods   C57BL/6 mice were randomly divided into blank control group (n=8), model group (n=12), sham electroacupuncture group (n=6) and electroacupuncture group (n=6). Except blank control group, mice in other groups were injected intraperitoneally with LPS (0.25mg/kg) for 7 consecutive days. On the 8th day, mice in the sham electroacupuncture group and electroacupuncture group were treated with acupuncture or Zusanli electroacupuncture for 7 consecutive days. The mice were weighed before the experiment, on the 7th and 14th days. On the 13th day, the elevated cross maze test was performed on the mice. The open field test was performed on the 14th day. After the experiment, immunofluorescence assay was used to determine the expression of microglial ionized calcium binding adaptor molecule-1(Iba-1) in prefrontal cortex region. The mRNA expression of NF-κB, inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), Caspase-1 and interleukin (IL)-18 were detected by Real-time PCR. The protein expression levels of NF-κB, iNOS, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, IL-1β and IL-18 were detected by Western blotting.  Results   Weight change, On the 7th day, compared with the control group, the body weight of mice in model group, sham electroacupuncture group and electroacupuncture group decreased (P<0.0001), respectively; On the 14th day, compared with the control group, the weight of mice in the model group decreased (P<0.0001); Compared with the sham electroacupuncture group, the body weight of mice in the electroacupuncture group increased (P<0.05). Elevated cross maze experiment, compared with the control group, the total distance and open arm retention time of mice in model group decreased, while the closed arm retention time increased (P<0.05). The open field experiment showed that compared with the control group, the model group mice showed a decrease in total distance traveled, slower movement speed, and fewer entries into the central area(P<0.001); Compared with the model group, the electroacupuncture group showed an increase in all three indicators(P<0.01); Compared with the sham electroacupuncture group, the total distance and motion speed of mice in electroacupuncture group both increased (P<0.05). Immunofluorescence assay, compared with the control group, the relative fluorescence of Iba-1 in prefrontal cortex area of mice in model group increased (P<0.05). Compared with the model and sham electroacupuncture group, the relative fluorescence of Iba-1 in prefrontal cortex area of mice in electroacupuncture group decreased (P<0.05). Real-time PCR showed that compared with the control group, mRNA expressions of NF-κB, iNOS, TNF-α, Caspase-1 and IL-18 in the model group increased (P<0.05); Compared with the model group, mRNA expressions of NF-κB, iNOS, TNF-α, Caspase-1 and IL-18 in electroacupuncture group decreased (P<0.05). Western blotting indicated that compared with the control group, the protein expressions of NF-κB, iNOS, Caspase-1, IL-1β and IL-18 in model group increased (P<0.05); Compared with model group, the protein expressions of NF-κB, iNOS, NLRP3, ASC, Caspase-1, IL-1β and IL-18 in electroacupuncture group decreased (P<0.05); Compared with the sham electroacupuncture group, IL-18 protein in electroacupuncture group decreased (P<0.05).  Conclusion   Electroacupuncture can improve the behavioral performance of mice and inhibit the activation of microglia in the cortical region of mice, which may play an anti-inflammatory and protective role by regulating NF-κB/ NLRP3 pathway. 
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    Cell and Molecules Biology
    Angelica Sinensis polysaccharide promoting hematopoietic reconstruction in receptor mice after bone marrow transplantation
    YANG Ting LIAO Kui HUANG Cai-hong WEI Han WANG Cheng DU Kun-hang WANG Zi-ling WANG Lu WANG Ya-ping
    2024, 55 (5):  556-564.  doi: 10.16098/j.issn.0529-1356.2024.05.006
    Abstract ( )   PDF (4465KB) ( )  
    Objective   To explore the mechanism of Angelica Sinensis polysaccharide (ASP) promoting donor bone marrow transplantation (BMT) to reconstruct hematopoietic function of receptor mice by regulating bone marrow stromalcells (BMSCs).  Methods   Bone marrow mononuclear cells (BMMNCs) of male C57BL/6J mice aged 8-10 weeks were separated, purified and transplanted into female receptor mice of the same age. On the ninth day, receptor mice BMMNCs were separated, purified and transplanted again into female receptor mice. The transplanted receptor mice were divided into control group:  sham irradiation; Irradiation(IR) group: a whole-body irradiation with a total dose of 8.0 Gy X-ray; BMT group: the receptor mice treated in the same way as the IR group and transplanted BMMNCs (5×106 cells) from male donor via the tail vein; BMT+ASP group: the receptor mice treated in the same way as the BMT group, and injected ASP [100 mg/(kg·d)×9] by intraperitoneal route from the first day of transplantation. Changes in body weight and survival rate of mice were recorded during modeling, receptor mice BMMNCs were collected to detect sexdetermining region of Y(SRY) gene after building model, peripheral blood indexes, the number of BMMNCs in femur and histopathology of bone marrow were detected; BMSCs in receptor mice was separated and purified, BMSCs adhesion ability was observed, proliferation ability was detected by 5-ethynyl-2-deoxyuridine(EdU);The level of reactive oxygen species (ROS), the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in BMSCs were detected; The levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), stem cell factor(SCF), insulinlike growth factor 1(IGF-1) in culture supernatant of BMSCs were determined, CFU-Mix was counted after BMMNCs co-cultured with receptor BMSCs in each group for 48 hours;The expression of Notch signaling pathway related genes (Notch1, Jagged1, Hes1) in BMMNCs were measured by Real-time PCR.    Results   All mice in IR group were died,the body weight loss in BMT+ASP group was not obvious. The SRY gene was detected in the receptor female mice BMMNCs. Peripheral blood indexes and the number of BMMNCs were not significantly decreased in BMT+ASP group receptor mice,and bone marrow histopathological injury was reduced. ASP promoted the proliferation of BMSCs, decreased the contents of ROS and MDA, and increased the activity of SOD in BMSCs. ASP promoted the secretion of SCF, GM-CSF and IGF-1 in BMSCs, and increased CFU-Mix yield of BMMNCs co-cultured with receptor BMSCs. ASP increased the expression of Notch1, Jagged1 and Hes1 mRNA in BMMNCs.  Conclusion   The mechanism of ASP promoting receptor hematopoietic function reconstruction is related to reducing the oxidative stress damage of hematopoietic microenvironment, improving the secretion of hematopoietic growth factors in BMSCs, and regulating Notch signaling pathway. 
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    Regulation of inflammatory factors in the coronary atherosclerosis region on stem cell migration in mice
    YANG Yun SUN Pan-wen XU Ya-ping GUO Zhi-kun
    2024, 55 (5):  565-572.  doi: 10.16098/j.issn.0529-1356.2024.05.007
    Abstract ( )   PDF (6494KB) ( )  
    Objective   To investigate the effects of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in coronary arteriosclerosis (AS) on the migration of stem cell antigen-1(Scal-1) and Nanog positive cells migration.  Methods   Ten atherosclerotic model mice and 10 normal mice were taken respectively, and the fresh hearts of the two groups were taken out, OCT embedding, frozen sections, HE staining and oil red O staining were performed respectively to observe the pathological changes of coronary arteries; Using immunohistochemical staining techniques, the expression changes of TNF-α, IL-6, Scal-1, and Nanog in the coronary artery wall were observed under light microscope. The ventricular tissue of 20 1-week-old mice was excised. Primary Scal-1 and Nanog positive cells were extracted using density gradient centrifugation, and these cells were purified and cultured. Immunofluorescence technology was used to identify the purity of Scal-1 and Nanog positive cells. Transwell migration experiments on passage 1(P1) cells was performed to observe the effects of different concentrations of inflammatory factors on the migration of Scal-1 and Nanog positive cells.  Results  The blood lipid test showed that the total cholesterol, triglycerides, and low-density lipoprotein cholesterol of the model mice were significantly higher than those of the normal control group. Histological staining showed that all the mouse models had obvious coronary atherosclerosis plaque. Immunohistochemical staining results showed that the inflammatory factor TNF-α and IL-6 in atherosclerosis region were strong positive expression, while Scal-1 and Nanog positive cells in atherosclerotic plaques increased, compared to the normal group. The positive rate of Scal-1 in P1 generation cells was about 80%, The positive rate of Nanog was approximately 91%. The Transwell experiment showed that IL-6 had no effect on cell migration at low concentrations, but had an inhibitory effect on cell migration at high concentrations; Low concentrations TNF-α promoted cell migration with 0.5 μg/L TNF-α showing the most significant effection cell migration, and exhibiting inhibitcry effects inhibit at high concentrations.  Conclusion   TNF-α  and IL-6 inflammatory factors, Scal-1 and Nanog positive cells are all involved in the occurrence and development of AS, and the appropriate concentration of inflammatory factors can enhance the migration of stem cells to the coronary atherosclerosis region. 
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    Cancer Biology
    Non-structural maintenance of chromosome condensin Ⅰ complex subunit G promoting the proliferation, migration and invasion of ovarian cancer cells by the Akt signaling pathway
    ZHANG Ming-shu WANG Yi-hui ZHANG Qing YE Li-ping
    2024, 55 (5):  573-581.  doi: 10.16098/j.issn.0529-1356.2024.05.008
    Abstract ( )   PDF (4692KB) ( )  
    Objective   To explore the regulatory mechanism of non-structural maintenance of chromosome condensin I complex submit G (NCAPG) on proliferation, migration, and invasion of ovarian cancer cells.  Methods   The bioinformatics database was used to analyze the differential expression of NCAPG in ovarian cancer tissues. Western blotting was used to detect the protein expression of NCAPG in normal ovarian epithelial cells IOSE80, ovarian cancer A2780 and SKOV3 cells. The silencing experiments of NCAPG siRNA were divided into blank, control, siNCAPG-1 and siNCAPG-2 groups. The overexpression experiments of NCAPG plasmid were divided into blank, control, NCAPG, NCAPG+MK2206 and MK2206 groups. MTT assay was used to detect cell proliferation activity. Cell scoring assay and transwell assay were used to analyze cell migration and invasion. The protein expressions of p-Akt, total(t)-Akt, proliferative cellular nucleus antigen (PCNA), matrix metallopeptidase 9 (MMP-9), vimentin, N-cadherin, and E-cadherin were detected by Western blotting.  Results   NCAPG was highly expressed in ovarian cancer tissues and cells. Knockdown of NCAPG significantly inhibited the proliferation, invasion and migration of ovarian cancer SKOV3 cells. The protein expressions of p-Akt, PCNA, MMP-9, vimentin and N-cadherin decreased while E-cadherin expression increased. Overexpression of NCAPG significantly promoted the proliferation, invasion and migration of ovarian cancer A2780 cells. The protein expressions of pAkt, PCNA, MMP-9, vimentin, and N-cadherin increased while E-cadherin expression decreased. Akt inhibitor MK2206 significantly attenuated the above effects of NCAPG.  Conclusion  NCAPG promotes the proliferation, invasion, and migration of ovarian cancer cells by activating the Akt signaling pathway and regulating the expression of PCNA, MMP-9, and epithelial-mesenchymal transition (EMT)-related proteins. 
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    Dipeptidylpeptidase-4 inhibitor anagliptin inhibiting the formation and mechanism of lung metastasis in colorectal cancer 
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    LI Yue WEI Si-meng WU Xin LIU Xiao LI Qi CHEN Chang
    2024, 55 (5):  582-588.  doi: 10.16098/j.issn.0529-1356.2024.05.009
    Abstract ( )   PDF (4980KB) ( )  
    Objective  To investigate the formation and mechanism of dipeptidylpeptidase-4 inhibitor anagliptin inhibiting lung metastasis of colorectal cancer.  Methods   Twenty-four male BALB/c mice were randomly divided into normal group, model group, anagliptin group. The control group did not undergo any treatment. The model group was injected with 0.1 ml CT-26 cells of 109 cells/L. once through the tail vein of the mouse to construct a lung metastasis model, while the anagliptin group was injected intraperitoneally 20mg/(kg·d) 0 day after  constructing a lung metastasis model. The mice were sacrificed after 14 days. HE staining was used to detect the morphological alteration. Determination of CT-26 cell viability through MTT assay and CT-26 cell apoptosis was detected by flow cytometry and live/dead cell staining. Reactive oxygen species(ROS) production was measured by ROS kit. Western blotting was conducted to measure the expression level of Bcl-2, Bax, cleaved-caspase-3, superoxide dismutase(SOD-1) and SOD-2.  Results   HE staining showed that the administration of anagliptin could significantly inhibit the abnormal changes in the model group. Anagliptin inhibited the viability of CT-26 cells above 2 mmol/L. Anagliptin promoted the apoptosis of CT-26 cells. Incubation of anagliptin in CT-26 cells significant promoted the production of ROS. Incubation of anagliptin stimulated the expressions of Bax and cleaved-Caspase-3, while down-regulated the expression of Bcl-2 in CT-26 cells. Administration of anagliptin decreased the expression of SOD-1, but not SOD-2.  Conclusion   Anagliptin promotes apoptosis of colorectal cancer cells and inhibits the formation of lung metastatic tumors through the SOD-1/ROS pathway.  
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    Anatomy
    Comparison on cone beam CT measurement of distal space of mandibular molars between average growth pattern’ s adult patients with skeletal class Ⅲ and skeletal class Ⅰ
    ZHAO Ying-bin FENG Jian-kun
    2024, 55 (5):  589-594.  doi: 10.16098/j.issn.0529-1356.2024.05.010
    Abstract ( )   PDF (2890KB) ( )  
    Objective  To measure the distal space between skeletal class Ⅲ and skeletal class Ⅰ mandibular molars, compare the differences, and to measure the contact situation between the distal root tip of the second mandibular molar and the lingual cortical bone of the mandible, providing a theoretical reference for the distance of mandibular molar distal movement by using cone beam CT(CBCT).  Methods   CBCT imaging data of 60 adult patients who received orthodontic treatment at Chengde Stomatological Hospital from 2020 to 2022 were selected, including 30 patients with skeletal class Ⅲ mean angle and 30 patients with skeletal class Ⅰ mean angle. CBCT image data were exported to DICOM format, then import it into Mimics 20.0 software, and measure the distal space of bilateral mandibular second molars. The contact between the distal root tip of the second mandibular molar and the lingual bone cortex of the mandible was measured. Import the measurement results into SPSS 19.0 statistical software to analyze the differences and the contact rate.  Results   There was no statistical difference in the measurement results of the distal space of mandibular molars between genders, and sides, and between mandibular molars with or without wisdom teeth (P>0.05). The distal space of mandibular molars in skeletal class Ⅲ were greater than those in skeletal class I, and the difference was statistically significant (P<0.05). The contact rate between the distal root tip of the second mandibular molar and the lingual bone cortex of the mandible was 34.83%. There was no significant difference in the contact rate between the distal root tip of the mandibular second molars and the lingual cortical bone of the mandible between class Ⅲ malocclusion and class I malocclusion(P>0.05).  Conclusion   There is a certain gap in the distal part of the skeletal class Ⅲ mandibular molar. In clinical practice, the molar can be designed to move distally, expanding the dental arch from the sagittal direction to provide a gap for the adduction of the mandibular anterior teeth. However, it is necessary to evaluate the contact between the distal root tip of the second mandibular molar and the lingual bone cortex of the mandible. If the distal root tip of the second mandibular molar contacts the lingual bone cortex of the mandible, the distal movement of the molars should be avoided. 
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    Histology,Embryology and Developmental Biology
    Regulatory effect of nobiletin on platelet-activating factor in diabetic rats with renal injury
    TONG Sen LUO Shi-cui YANG Qiu-qiong SONG Bo YANG Yu-qing WU Jun-zi
    2024, 55 (5):  595-603.  doi: 10.16098/j.issn.0529-1356.2024.05.011
    Abstract ( )   PDF (7948KB) ( )  
    Objective  To investigate the effect of nobiletin on plateletactivating factor (PAF) metabolism in diabetic rats with renal injury.  Methods   Totally 72 rats were randomly divided into control group (n=10) and modeling group (n=62). The modeling group rats were induced to develop a diabetic rat model with renal injury and then further divided into the model group, aspirin group (20mg/kg), and nobiletin low(50mg/kg), medium(100mg/kg), and high-dose (200mg/kg) groups, each with 10 rats. After continuous oral administration for 6 weeks, rat body weight, kidney weight, and kidney index were measured. Histopathological assessments were conducted by using HE, periodic acid-Schiff staining (PAS), Masson staining, and transmission electron microscopy. Blood glucose levels, renal function, inflammatory factors, PAF and its regulatory factors were detected. Expression levels of PAF metabolism-related proteins, PAF-acetylhydrolase(PAFAH), PAF receptor (PAFR), and cholinephosphotransferase 1(CHPT1) in kidney tissues were assessed using Western blotting and immunohistochemistry.  Results   Following nobiletin intervention, rat body weight increased while kidney weight and kidney index decreased. Improvement in renal tissue pathology was observed, with reduced interstitial fibrosis and thinner basement membrane. Fasting blood glucose and glycated hemoglobin decreased, while fasting insulin showed no significant improvement. Urea nitrogen, blood creatinine, cystatin C, and 24-hour urinary protein excretion were reduced. Levels of interleukin (IL)-1α, IL-6, IL-8, and tumor necrosis factor (TNF-α) were lowered. PAF and its regulatory factors decreased. PAFR and CHPT1 expression decreased, while PAFAH increased.  Conclusion   Nobiletin can alleviate renal injury in diabetic rats with renal injury, improve kidney function, regulate blood glucose, and mitigate inflammatory response. Its mechanism may be associated with the modulation of platelet-activating factor metabolism. 
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    Effect and mechanism of reactive oxygen species/p38 mitogen-activated protein kinase cascade on renal stone formation in rats
    XIE Ya-bin WANG Fei WANG Kang-yang LIN Shi-shuai
    2024, 55 (5):  604-611.  doi: 10.16098/j.issn.0529-1356.2024.05.012
    Abstract ( )   PDF (4045KB) ( )  
    Objective  To study the effect of reactive oxygen species (ROS)/p38 MAPK cascade reaction on the formation of kidney stones (KS) in rats and explore the mechanism.  Methods   Fifty SD rats were randomly divided into control group (normal feeding), N-acetylcysteine (NAC) group (intraperitoneally injected 200 mg/kg NAC), KS group (constructed calcium oxalate KS model), KS+NAC group (constructed calcium oxalate KS model, intraperitoneally injected 200 mg/kg NAC), KS+NAC+tunicamycin (TM) group(constructed calcium oxalate KS model, intraperitoneally injected 200 mg/kg NAC and 1 mg/kg TM), with 10 rats in each group. After 4 weeks of administration, 24 hours urine volume and oxalic acid (Ox) of each group were measured, serum creatinine (Cr), urea nitrogen (BUN) and uric acid (UA) levels were detected by automatic biochemical analyzer. HE staining and Von Kossa staining were used to observe the histopathological changes and crystal deposition of the kidney. TUNEL staining was used to detect apoptosis of renal tissue cells. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in renal tissue were measured by the kit. DHE fluorescent probes detected the levels of reactive oxygen species (ROS) in kidney tissue. Immunohistochemical staining was used to detect the expressions of microtubule-associated protein 1 light chain 3 B (LC3B) and glucose regulatory protein 78 (GRP78) in renal tissue, and the protein expression of LC3Ⅱ/LC3Ⅰ, Beclin1, GRP78, CCAAT/enhancer binding protein homologous protein (CHOP) in renal tissue was determined by Western blotting.  Results   Compared with the KS group, Ox in KS+NAC group decreased (P<0.05), BUN, Cr and UA levels decreased (P<0.05), renal tubule dilatation and calcium oxalate crystallization decreased, TUNEL positive cell rate decreased (P<0.05), SOD activity increased and MDA content decreased (P<0.05), ROS levels decreased (P<0.05), LC3B and GRP78 positive staining levels decreased (P<0.05), the relative protein expressions of p-p38 MAPK/p38 MAPK, LC3Ⅱ/LC3Ⅰ, Beclin1, GRP78 and CHOP were down-regulated(P<0.05). Compared with the KS+NAC group, Ox in KS+NAC+TM group increased (P<0.05), BUN, Cr and UA levels also increased (P<0.05), renal tubule dilated significantly, calcium oxalate crystals increased, TUNEL positive cell rate increased (P<0.05), SOD activity decreased and MDA content increased (P<0.05), ROS levels increased (P<0.05), LC3B and GRP78 positive staining levels increased (P<0.05), the relative protein expressions of p-p38 MAPK/p38 MAPK, LC3Ⅱ/LC3Ⅰ, Beclin1, GRP78 and CHOP were also up-regulated (P<0.05).  Conclusion   ROS/p38 MAPK cascade is involved in promoting KS formation in rats, which is related to the activation of endoplasmic reticulum stress-mediated autophagy pathway. 
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    Distribution and localization of dopamine receptor in small intestines
    CHEN Jun-jun ZHOU Li SU Tian WANG Xian-wei ZHANG Hai-long WANG Zhi-yong
    2024, 55 (5):  612-618.  doi: 10.16098/j.issn.0529-1356.2024.05.013
    Abstract ( )   PDF (8951KB) ( )  
    Objective  To explore the distribution and localization of dopamine receptor D3-D5 in the small intestine of different species.  Methods   The distribution and expression of D3-D5 in the small intestine of mice, rats and rhesus monkeys were detected by immunohistochemistry and Western blotting. The expression of D3-D5 in immunoglobulin A positive plasma cells (IgA+PC) located in the lamina propria (LP) were detected by immunofluorescence double labeling.  Results   D3 and D5 were widely distributed in the epithelium, LP, submucosal plexus (SMP) and intermuscular plexus (MP) of the small intestine in mice, rats and rhesus monkeys. The distribution of D4 in the small intestinal of mice and rhesus monkeys were consistent with the result  of D3 and D5. D4 was distributed only within the epithelium and LP of rat small intestine. D3 and D5 were expressed in the IgA+PC in the LP of mice and rats, whereas D4 was not.  Conclusion  The distribution and localization pattern of D3 and D5 are similar in the small intestine of mice, rats and rhesus monkeys, whereas those of D4 vary between different species. Dopamine may be involved in regulating the functions of IgA+PC. 
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    Anthropology
    General characteristics of Chinese ethnic groups based on body index value
    LI Yong-lan YU Hui-xin YU Ke-li ZHANG Xing-hua BAO Jin-ping ZHENG Lian-bin
    2024, 55 (5):  619-624.  doi: 10.16098/j.issn.0529-1356.2024.05.014
    Abstract ( )   PDF (1829KB) ( )  
    Objective  To explore the common features of Chinese ethnic groups.  Methods   Eight body indexes of 62 ethnic groups in China were analyzed.  Results   The cluster analysis showed that 52 males and 59 females ethnic groups were grouped into the mixed group dominated by the northern ethnic group and the mixed group dominated by the southern ethnic group. Eight Han ethnic groups were grouped into each group, but no Han group was aggregated. The result  of body index classification showed that the main body types of Chinese male population were long trunk, middle chest, wide shoulder, wide pelvis and middle leg. Middle body, wide chest, wide shoulder, wide pelvis and middle leg were the main body types of Chinese female population. This showed that the characteristics of Chinese ethnic groups had obvious consistency. The consistency of Chinese group features was related to its close origin. It should be said that Han nationality played an important role in the process of communication and integration of various ethnic groups in China. In the history of the Han nationality, there had been many large-scale population migration. The southern movement of the northern ethnic minorities into the northern Han and the southward movement of the northern Han into the south promoted the formation of the Southern Han, which made the southern Han and the northern Han had similar body features, and also promoted the southern ethnic minorities into the southern Han. In addition, the Han nationality who moved into minority areas also gradually integrated into minority areas. Conclusion   There are obvious commonalities in Chinese ethnic groups. 
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    Cephalometric parameters of three Wa dialect ethnic groups in China
    YAO Yue-tong YU Ke-li ZHANG Xing-hua GAO Xin-ying XIAO Yao CHENG Zhi GAO Wen-fang LIU Xin BAO Jin-ping
    2024, 55 (5):  625-631.  doi: 10.16098/j.issn.0529-1356.2024.05.015
    Abstract ( )   PDF (1563KB) ( )  
    Objective  To survey and analysis of cephalometric indicators of Wa adults in China.  Methods  Cephalometric parameters were measured in 1996 cases (858 males and 1138 females) of Wa adults in China, including 927 cases (381 males and 546 females) of the Baraoke ethnic group, 564 cases (241 males and 323 females)of the A Wa ethnic group, and 505 cases (236 males and 269 females) of the Wa ethnic group by using sliding caliper and spreading caliper. Seventeen direct cephalofacial parameters and one indirect parameter for each of the three dialect ethnic groups were derived separately and analyzed for age correlations, inter-sex u-tests,and multiple comparisons. Finally, the three dialect ethnic groups were subjected to cluster analysis and principal component analysis with 15 ethnic groups in China.  Results   Nose breadth, mouth breadth and physiognomic ear length were significantly and positively correlated with age for both sexes in the three Wa dialect ethnic groups, while head breadth and lip height were significantly and negatively correlated with age. Except for the interocular breadth, there were gender differences between males and females in the cephalometric parameters of the three Wa dialect ethnic groups. The cephalofacial features of the Baraoke, A Wa and Wa ethnic groups were different, as evidenced by the fact that males and females of the Baraoke and Wa dialect ethnic group had higher lip height, wider nasal breadth and wider mouth breadth, while males and females of the A Wa ethnic group had lower nasal height.  Conclusion   The cephalofacial features of the three Wa dialect ethnic groups are close to those of the Khmus and Mang, who have their origins in the ancient Baipu people and are also members of the Mon-Khmer language group of the Austroasiatic linguistic. 

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    Bioengineering
    Evaluation of biological properties of Gd-doped hydroxyapatite bio-nanocomposites
    KONG Wei-li YANG Yu SHEN Fu-guo SUN Wen-cai GU Hao JIN Song XIAO Wen-long
    2024, 55 (5):  632-640.  doi: 10.16098/j.issn.0529-1356.2024.05.016
    Abstract ( )   PDF (6554KB) ( )  
    Objective  To investigate the biocompatibility of new gadolinium-doped hydroxyapatite (Gd-HA) composite scaffolds and to explore their feasibility as cell culture materials and bone tissue engineering scaffolds.Methods  The Gd-HA composite scaffolds were chemically synthesized and placed under the electron microscope for observation. The experiment was divided into three groups, the HA group, the Gd-HA group, and the control group.Rabbit adipose-derived mesenchymal stem cells (ADSCs) were isolated, cultured and characterized, and the Gd-HA composite scaffold extract was added to the ADSCs in vitro culture system. Cell survival and cytotoxicity were assessed by live-dead cell staining, cell proliferation ability within the scaffolds was assessed by CCK-8 assay, and the scaffolds were assessed by alizarin red staining for cell osteogenic differentiation. The toxic reactions of the scaffold materials were observed by skin irritation test, systemic acute toxicity test and muscle tissue and liver and kidney pathology at the site of intramuscular implantation of the scaffolds.  Results  The Gd-HA composite scaffold showed irregular void structure under electron microscope. Cell morphology observation showed that ADSCs grew adherently to the wall and were long shuttle-shaped. The positivity rate of CD29 was 96.94%, CD44 was 97.90%, CD45 was 0.10%, and CD34 was 0.46%, which was obtained using flow cytometry. Live-dead cell staining showed that the amount of live cells in the Gd-HA group was significantly better than that in the hydroxyapatite(HA) group after 5 days of co-culture. CCK-8 assay showed no significant difference in cell proliferation within 0-3days. After 3days, the Gd-HA group was significantly better than the HA group and the control group (P<0.05). Calcium nodule deposition after alizarin red staining was significantly better in the Gd-HA group than in the HA and control groups, showing a deeper red color. No skin irritation was observed in gross and skin tissue HE observations after the contact of the extract with the skin. The general condition of the experimental groups was good after the infusion of the extract into the abdominal cavity, and the body mass tended to increase steadily (P>0.05). HE staining showed that inflammatory reaction at the interface between the material and muscle tissue of the stent intramuscular implantation site in Gd-HA group was significantly higher than that of the control group, and the inflammatory cell infiltration was gradually reduced with the prolongation of implantation time. At the 8th weeks the morphology of the tissue around the material was close to normal muscle tissue, and no pathological changes were observed in the HE staining of liver and kidney at the 12th week. Conclusion   Gd-HA composite scaffolds exhibit good biocompatibility and facilitate cell proliferation and osteogenic differentiation, and they are expected to serve as good carriers for stem cell transplantation in tissue engineering. 
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    Review
    Current progress of metal ions in the pathogenesis of refractory epilepsy 
    LI Hao ZHANG Ying LIU Yuan-yuan
    2024, 55 (5):  641-646.  doi: 10.16098/j.issn.0529-1356.2024.05.017
    Abstract ( )   PDF (1085KB) ( )  
    Epilepsy is a common chronic neurological disorder caused by abnormal electrical activity in the brain, and its pathogenesis has not been fully clarified. In recent years, more and more studies have shown that iron, zinc and copper in the brain are involved in the occurrence and development of epilepsy. These metal ions possibly play a role in epileptogenesis by affecting neuronal excitability and signal transduction through alterating ion channel function, potential balance, action potential propagation, redox and so on. In this review, we mainly analyse the effects of these metal ions on epilepsy, in order to explore the underlying mechanisms of epilepsy and provide promising therapeutic strategies for epilepsy based on metal ions. 
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    Advances in research on neuromodulation of ovarian cancer
    LI Fan-shu SUN Yan-rong ZHANG Yan LUAN Li-ju ZHANG Wei-guang
    2024, 55 (5):  647-653.  doi: 10.16098/j.issn.0529-1356.2024.05.018
    Abstract ( )   PDF (1344KB) ( )  
    Ovarian cancer is one of the most common gynecologic cancers in the world. Over the past few decades, there has been considerable research reporting on the mechanisms of cancer development and progression, with multiple nerve as well as neurotransmitters involved. Nerve innervation is also found in ovarian cancer. And in ovarian cancer, various nerves and neurotransmitters play different roles. They are involved in ovarian cancer cells’ proliferation metastasis, apoptosis and changes in the tumor microenvironment. Further understanding of the role of these nerve endings in the development of ovarian cancer is essential for understanding the mechanisms of cancer progression. This will be important for subsequent research focusing on tumor regulation. While glucocorticoids and sympathetic nerve-released norepinephrine are able to promote ovarian cancer progression, serotonin may inhibit cancer cell growth. Also, parasympathetic and sensory nerves are capable of having either a positive or negative effect on ovarian tumors. These relevant studies offer the possibility of new therapeutic options for oncology, it may be possible to mitigate the progression of cancer with inexpensive receptor inhibitors or agonists. This will facilitate the subsequent exploration of therapeutic possibilities forovarian cancer and other cancer-related treatments. In this review, we also present some insights into the role of the nervous system in the regulation of ovarian cancer, which we hope will provide new insights into the innervation and progression of ovarian cancer. 
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