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    Technology and Methodology
    Complete Box Fusion based on Ensemble Networks for rib fracture detection and localization
    HE Xue-cai JIN Liang LI Ming ZHANG Chen-xi
    2022, 53 (3):  396-401.  doi: 10.16098/j.issn.0529-1356.2022.03.019
    Abstract ( )   PDF (1607KB) ( )  
    Objective  To propose a new rib fracture detection network Rib-Net to automatically and accurately detect and locate rib fracture and address the issue of missed diagnosis of rib fractures.   Methods  The public data set RibFrac Dataset was used to evaluate the performance of the Rib-Net, and the data set was divided into training set (420 cases), validation set (80 cases), and test set (160 cases). The Rib-Net was composed of the object detection integrated network Ensemble Detection Net (ED-Ne), Complete Box Fusion (CBF) module and the segmentation network 3D Unet. Firstly, Retina Unet, UFRCNN+ and Mask RCNN were integrated to form ED-Net to predict rib fracture candidate boxes. Secondly, a new CBF module was designed to fuse overlapping fracture candidate boxes to generate candidate boxes with accurate positioning and accurate confidence. Finally, Unet was used for rib fracture segmentation to achieve further precise localization of rib fractures.   Results  On the “MICCAI 2020 RibFrac Challenge: Rib Fracture Detection and Classification challenge”, our proposed Rib-Net’s detection results  reached the best performance, and its recall rate, free-response  receiver operating characteristic curve(FROC) value and Dice were 92.3%, 0.859 and 0.61, respectively.   Conclusion  The Rib-Net network can efficiently and accurately detect and locate rib fractures on chest CT images, effectively assisting doctors in making accurate diagnosis. 
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    Modified induction method for differentiation of rat pancreatic ductal stem cells to form islet-like cells
    ZHANG Wen-qi YANG Jie WU Jiang LANG Guan-cun XIAO Mei
    2022, 53 (3):  387-395.  doi: 10.16098/j.issn.0529-1356.2022.03.018
    Abstract ( )   PDF (7518KB) ( )  
    Objective  To establish a modified induction method  for differentiation of rat pancreatic ductal stem cells (rPDSCs) to form islet-like cells.   Methods  All-trans retinic acid(ATRA) was added at 2, 4, 6 and 8 μmol/L in the basal culture medium DMEM/F12 + 10% FBS + 1% penicillin/1% streptomycin to induce the differentiation of rPDSCs to form islet-like cells in vitro, and the optimal induction concentration of ATRA was screened. Based on the optimal ATRA induction concentration, rPDSCs were then induced to form islet-like cells in vitro by matrigel culture, suspension culture or hanging drop culture, respectively, to screen the optimal induction culture method . Cell morphology, dithizone(DTZ) staining, cell immunofluorescence staining, Real-time PCR and ELISA were used to detect the induced islet-like cells.  Results  Compared with the control group, 6 μmol/L ATRA and matrigel culture were the best in the basic culture medium. After 28 days of induction, the cells enriched and differentiated to form islet-like spherical cell clusters; DTZ staining was positive; Pancreatic duodenal homeobox-1(Pdx1) and insulin were expressed at gene and protein levels, respectively; Glucose stimulation, release insulin and C-peptide, showed glucose concentration dependent.   Conclusion  The in vitro differentiation of rPDSCs to form islet-like cells could be successfully induced by using 6 μmol/L ATRA+DMEM/F12+10% FBS+1% double antibody under matrigel culture method in the present study.
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    Anthropology
    Association between methylenetetrahydrofolate reductase polymorphisms and hypertensive disorders of pregnancy in Han nationality in Qinghai Province
    DUAN Qian LI Jian-hua LI Wen-jia WANG Ru
    2022, 53 (3):  354-359.  doi: 10.16098/j.issn.0529-1356.2022.03.013
    Abstract ( )   PDF (1285KB) ( )  
    Objective  To investigate the association between polymorphisms of methylene tetrahydrofolate reductase (MTHFR)-677C/T (rs1801133) and hypertensive disorders of pregnancy (HDP) in Han women of Qinghai Province.   Methods  A total of 139 HDP subjects (HDP group) and 145 normal pregnant subjects (control group) were selected from Qinghai province. The MTHFR-677C/T polymorphism typing in HDP group and control was analyzed by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).The mutation was confirmed by sequencing.  Results  The frequency of CC CT TT genotype of MTHFR gene in HDP group and control group were 54.68%,35.25%,10.07% and 69.66%,22.06%,8.28% respectively (P<0.05);The frequency of CC genotype (54.68%) in HDP group was lower than that(69.66%) in control group (P<0.05),the frequency of CT genotype ( 35.25%) in HDP group was higher than that (22.06%) in control group (P<0.05),there was no significant difference in the frequency of TT genotype between HDP group and control group (P>0.05);The frequency distribution of alleles C and T of MTHFR-677C/T polymorphisms were different between HDP group and control group (P<0.05),T allele frequency in HDP group was higher than that in control group(χ2=5.568,P<0.05).   Conclusion  The polymorphisms of MTHFR -677C/T is significantly correlated with hypertensive disorders of pregnancy of Han group in Qinghai Province. The T allele of MTHFR -677C/T polymorphism may be the susceptibility gene of HDP,CT genotype is the susceptibility genotype of HDP.
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    Phenotypic characteristics of head and face of Tajik adults in Aketao
    LI Qiang LI Xin MAIERDANJIANG Mohe-taer ALIMIRE Aier-ken MEIHERIGULI Wujimaimai-ti MAHERE Tuerhong MAIREMUGULI Kuer-ban ZHANG Wen-qian WEN You-feng
    2022, 53 (3):  360-366.  doi: 10.16098/j.issn.0529-1356.2022.03.014
    Abstract ( )   PDF (1319KB) ( )  
    Objective  To obtain the physical anthropological characteristics of head and face of Tajik adults in Aketao, and to explore their ethnic origin through cluster analysis with other populations.   Methods  According to Anthropometric Method, 286 Tajik adults (male 121, female 165)in Aketao, Xinjiang were recruited and analyzed. The survey included 12 observation indicators of head and face, 19 measuring indicators of head and face, 12 indexes of head and face and its classification.   Results  The indicators of head length, head breadth, head height, minimum frontal breadth, face breadth, bigonial breadth, nose breadth, mouth breadth, auricular height, physiognomic facial height, morphological facial height, nasal length, nasal height, physiognomic ear length, physiognomic ear breadth, zygomatic protection, nasal root height, nasal profile, nasal base of Tajik adults in Aketao had gender differences(P<0.05). Compared with the Tajik adults in Aketao and Taxkorgan, the type indicators of head and face of zygomatic protection, direction of eyeslits, nasal root height, nasal base, maximal diameter of nostrils, lobe types, nreadth-height index of head type, morphological facial index type, nasal index type had regional differences(P<0.05). The cluster analysis showed that Tajik adults have unique characteristics of head and face, which were grouped separately and were more similar to Iranians.   Conclusion  Tajik adults in Aketao have the characteristics of head and face of the West Eurasian. The characteristics of head and face of Tajik adults in Aketao and Taxkorgan are somewhat different. From characteristics of head and face, they may be related to the Eastern Iranian tribe. 
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    Body characteristics of the Zhuang-Dong ethnic groups in China
    YI La-na LI Yong-lan YU Hui-xin
    2022, 53 (3):  367-373.  doi: 10.16098/j.issn.0529-1356.2022.03.015
    Abstract ( )   PDF (1420KB) ( )  
    Objective To investigate the body characteristics of zhuang-dong ethnic group.   Methods  Totally 14 Zhuang-Dong languages in Guizhou, Guangxi, Yunnan, Hainan and Hunan were measured with anthropometry.  Results The foot breadth, upper arm length and forearm length of male of Zhuang-Dong ethnic group were not correlated with age, while crista iliaca breadth was positively correlated with age (P<0.01), 24 indexes including weight, stature and tragion height are negatively correlated with age (P<0.05 or P<0.01). Six indicators of Zhuang-Dong female’s foot breadth, hand length, height of suprasternal notch above sitting plane, upper extremity length, lower extremity length and upper arm length were not related with age, while four indicators of hand breadth at metacarpale, crista iliaca breadth, total arm length and forearm length were positively correlated with age (P<0.05 or P<0.01), while 18 indicators of weight, stature and tragion height were negatively correlated with age (P<0.05 or P <0.01). The vast majority of body index values had statistical significance among age groups. The average value of all body indexes of Zhuang-Dong ethnic groups was larger than that in female (P<0.05 or P<0.01), that was,   he height, length and width of male body were larger than that of female. Principal component analysis showed that, the stature, sitting height, height of suprasternal notch above sitting plane and weight of Lingao people, Li nationality men and women were larger than those of other 12 Zhuang-Dong language group. Mulao and Bajia had similar human body characteristics. In addition, the mean values of human body indexes of the 14 Zhuang-Dong language group, Bulang ethnic group and Gejia ethnic group were lower than those of the northern Han nationality, the Mongolian Balhu ethnic group, the Mongolian Erdos ethnic group, the Heshuo ethnic group in Qinghai, the Uzbek ethnic group, the southern Han nationality, the Mosuo ethnic group, the Qiang ethnic group and the Yi ethnic group.   Conclusion  Men and women in Zhuang-Dong ethnic group have little weight, while men’s height belongs to short stature and female’s height belongs to sub-medium stature. Zhuang-Dong ethnic group has the physical characteristics of ethnic minorities in southern China, and belongs to the physical type of short stature and light weight.
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    Three obesity indicators predicting central obesity in Naxi and Pumi ethnic groups in Yunnan Province and inter-ethnic comparison
    ZHANG Xin WANG Xu-jiao LI Jing MA Wei FAN Ning XU Fei
    2022, 53 (3):  374-380.  doi: 10.16098/j.issn.0529-1356.2022.03.016
    Abstract ( )   PDF (1779KB) ( )  
    Objective  To compare the predicted value of three different obesity indicators  of on center obesity of Naxi and Pumi adult  in Lijiang City, Yunnan Province; To calculate the best cut-off point of waist-to-height ratio(WHtR), body mass index(BMI) and abdominal volume index(AVI); And to compare the differences of BMI and WHtR between the Naxi, Pumi and North-South ethnic groups, so as to provide scientific reference for central obesity and related diseases.   Methods  By random sampling, 658 cases of Naxi adults (278 cases for men and 380 cases for women) and 496 cases of Pumi adults (216 cases for men and 280 cases for women) were selected as the subjects of the study,taking the internationally recognized visceral fat level(VFL) as the diagnostic criterion of central obesity, the relationship between WHtR, BMI, AVI and VFL was discussed by regression analysis, and the value of three obesity indicators was predicted by comparing the receiver operating characteristic(ROC) curve of the subjects. Using Excel 2019 to draw diagrams and SPSS 23.0 to process data.   Results  The six indicators of age,weight,height,waist circumference,hip circumference and AVI of Naxi and Pumi men were statistically significant among different ethnic groups (P<0.05); Naxi and Pumi women’s age, height, waist circumference, WHtR, BMI and abdominal volume index,the six indicators were statistically significant among different ethnic groups (P<0.01). Correlation analysis showed that the three indicators of WHtR, BMI and AVI were positively related to VFL (P<0.01).The ROC showed that the WHtR had the highest value for predicting central obesity in Naxi and Pumi men (P<0.01), BMI had the highest value of predicting central obesity in Naxi and Pumi women (P<0.01). Cluster analysis showed that Naxi and Pumi males were similar to southern ethnic groups such as Lisu and Sui, Naxi females were similar to Zang and Jingpo, and Pumi females were similar to southern ethnic groups such as Lisu and Sui.   Conclusion  WHtR, BMI, AVI to determine central obesity are statistically significant, can be combined with BMI and AVI to determine Naxi, Pumi central obesity.
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    Relationship between obesity indexes and body fat percentage in Naxi adults
    WANG Xu-jiao LI Jing ZHANG Xin MA Wei FAN Ning XU Fei
    2022, 53 (3):  381-386.  doi: 10.16098/j.issn.0529-1356.2022.03.017
    Abstract ( )   PDF (1369KB) ( )  
    Objective  To explore the relationship between obesity indexes and body fat rate in Naxi adults.   Methods  A total of 687 Naxi adults aged between 18 and 90 were selected from Yulong County, Lijiang City, Yunnan Province. Body weight, height, chest circumference, waist circumference, hip circumference, biceps skinfold, triceps skinfold, subscapular skinfold, suprailiac skinfold, anterior superior spine skinfold, body mass index(BMI),visceral fat index and body fat percentage were measured by anthropometry and bioelectrical impedance analysis. And the indexes and body fat percentage were statistically analyzed.   Results  The average body fat percentage of Naxi men was normal, while the women was obese. According to the criteria of visceral fat index, the average visceral fat index of Naxi men and women was within the normal range. According to the waist circumference, the waist circumference of Naxi men was within the normal range, while the waist circumference of women belonged to abdominal obesity. Correlation analysis showed that body weight, chest circumference, waist circumference, hip circumference, biceps skinfold, triceps skinfold, subscapular skinfold, suprailiac skinfold, anterior superior iliac spine skinfold, BMI, visceral fat index were positively correlated with body fat percentage among Naxi adults. There was the strongest correlation between visceral fat index and body fat percentage in Naxi men, followed by BMI; Naxi women’s chest circumference and body fat percentage, followed by waist circumference.   Conclusion  All the obesity indexes of Naxi adults are positively correlated with body fat percentage, and there are gender, regional and ethnic differences in the degree of correlation.
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    Review
    Advances in morphological and functional studies on the paraventricular thalamic nucleus
    LI Jia-ni LI Hui DONG Yu-lin LI Yun-qing
    2022, 53 (3):  402-406.  doi: 10.16098/j.issn.0529-1356.2022.03.020
    Abstract ( )   PDF (1531KB) ( )  
    Paraventricular thalamic nucleus(PVT)is an essential component of the midline thalamus, which has been regarded as a transmit relay nucleus and an integrated center in multiple behaviors including wakefulness, food intake, addiction, reward and fear memory. PVT is predominantly populated with glutaminergic excitatory neurons expressing vesicular glutamate transporter-2 (VGluT2) but without GABAergic inhibitory neurons. Therefore, based on the paradox of its multiplexed roles in different behaviors and its comparatively simplex excitatory nature, more specific subclassification of excitatory PVT neurons is required in studies in this field. In the present review, morphological and electrophysiological characteristics, efferent and afferent connections, and morphological and functional distinctions in anterior subregion and posterior subregion of PVT are summarized. In addition, neural connections and neurochemical properties are used as subclassification criteria in PVT neurons. This review might explain the integrated role of PVT in different behaviors, which would be helpful for further studies on the PVT. 
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    Histology,Embryology and Developmental Biology
    Expression of neuropeptide Y, calcitonin generelated peptide and inducible nitric oxide synthase in myocardium of rats with heart failure
    HUA Lei NING Shu-wei WANG Qian MENG Xiang-guang GUO Zhi-kun
    2022, 53 (3):  340-346.  doi: 10.16098/j.issn.0529-1356.2022.03.011
    Abstract ( )   PDF (6852KB) ( )  
    Objective  To explore the relationship between the content of neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), inducible nitric oxide synthase (iNOS) and the changes of the corresponding receptors in myocardial tissue and the development of heart failure.   Methods  Doxorubicin was used to prepare rat heart failure models(10 rats),control group (10 rats), Cardiac ultrasound testing evaluated its cardiac function, the distribution of nerve fibers was detected by nerve staining kit method, the tissue expression levels of NPY, CGRP, iNOS and neuropeptide Y receptor Y1 (NPY1R), receptor activity modifying protein 1 (RAMP1), guanylate cyclase beta 1(GCYB1) were observed by immunofluorescence and immunohistochemical staining method . Western blotting was used to detect the protein expression levels of NPY, CGRP, iNOS and NPY1R, RAMP1, GCYB1 receptors in myocardial tissue.   Results  Compared with the control group, the protein expression level and tissue expression distribution of CGRP in the myocardial tissue of the heart failure group were significantly decreased (P<0.05), while the density of nerve fibers,the protein expression level and tissue expression distribution of NPY and iNOS were significantly increased (P<0.05). The protein expression levels of GCYB1 and RAMP1 in myocardial tissue of the heart failure group were significantly increased, while the protein expression levels of NPY1R were significantly decreased (P<0.05).    Conclusion  Heart failure can lead to remodeling of the content and distribution of nerve fibers, CGRP, NPY, iNOS and their receptors in myocardial tissue, which may be one of the causes of cardiac innervation disorders.
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    Protective effects of micro RNA-98-5p targeting Kruppel-like factor 9 against myocardial ischemia-reperfusion injury in rats
    LU Fen YANG Qing SHAO Wen-jun SUN Zhen-kun WANG En-man ZHANG Wei-na
    2022, 53 (3):  347-353.  doi: 10.16098/j.issn.0529-1356.2022.03.012
    Abstract ( )   PDF (4380KB) ( )  
    Objective  To investigate the protective effect of micro RNA(miR)-98-5p targeting Kruppel-like factor 9(KLF9) against myocardial ischemia-reperfusion(MI/R) injury in rats.   Methods  Totally 50 rats were randomly divided into sham operation group, model group, miR-98-5p agomir group, agomir-NC group, and miR-98-5p agomir+pcDNA-3. 1-KLF9 group, 10 in each group. MI/R model was established by coronary artery ligation. The pathological changes of myocardial tissue were detected by HE staining. The myocardial apoptosis were detected by TUNEL. The levels of creatine kinase (CK), creatine kinase isoenzymes (CK-MB) and lactate dehydrogenase (LDH) in serum were detected by ELISA. The expression levels of miR-98-5p and KLF9 mRNA were detected by Real-time PCR. The expression of KLF9, Bax and JAK2/STAT3 pathway relative protein were detected by Western blotting. Dual luciferase assay verified the relationship between miR-98-5p and KLF9.  Results  Compared with the sham operation group, the arrangement of myocardial cells in the model group was disordered, and the myocardial cells appeared necrosis. The apoptosis rate of myocardial cells, serum CK, CK-MB and LDH contents increased, the expression level of miR-98-5p decreased, and the expression levels of KLF9 mRNA and protein, p-JAK2 and p-STAT3 protein increased (P<0.05). After the overexpression of miR-98-5p, the myocardial cells arranged more orderly and the myocardial cell necrosis decreased. The apoptosis rate of myocardial tissue, the contents of CK, CK-MB and LDH in serum and the expression of p-JAK2 and p-STAT3 protein were decreased (P<0.05). The result  of dual luciferase assay showed that KLF9 was the target gene of miR-98-5p. The overexpression of KLF9 reversed the effects of miR-98-5p agomir on myocardial injury.  Conclusion  MiR-98-5p targeting KLF9 can improve the myocardial injury of MI/R rats. The mechanism may be related to the regulation of JAK2/STAT3 signal pathway by miR-98-5p which inhibit myocardial cell apoptosis.
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    Neurobiology
    Dajianzhong decoction in treating visceral pain of irritable bowel syndrome based on ERK 1/2/nuclear factor κB signal pathway
    WU Jing YANG Sha-sha YANG Yi LI Yao-feng WANG Jun-xia CHEN Xiang-yun
    2022, 53 (3):  281-287.  doi: 10.16098/j.issn.0529-1356.2022.03.002
    Abstract ( )   PDF (2533KB) ( )  
    Objective To investigate whether Dajianzhong decoction can treat visceral pain of irritable bowel syndrome (IBS) by interfering with ERK1/2/nuclear factor κB (NF-κB )pathway and its downstream molecules.  Methods  The IBS visceral pain rat model was prepared by method  such as mother-infant separation, acetic acid enema, and intraperitoneal injection of chicken ovalbumin. Rats were randomly divided into normal control group (normal saline), model group (normal saline), Dajianzhong decoction (10.8g/kg) treatment group, and the control group of pinaverium bromide (45 mg/kg). Eight rats in each group were given intragastrically for 14 days. To evaluate the visceral sensitivity of rats, abdominal withdrawal reaction (AWR) was used; the expression of ERK1/2 mRNA, NF-κB mRNA, cyclooxygenase-2 (COX-2 )mRNA, and matrix metalloproteinase 9 (MMP-9)  mRNA in colon tissue of rats in each group were detected by Real-time PCR; Immunohistochemistry staining method  was used to detect the expression of NF-κB and COX-2 in the colon tissue of each group.   Results  Compared with the normal rats, AWR scores of model rats increased significantly at 60, 40 and 20 mmHg pressure (P<0.05, P<0.01), ERK1/2, NF-κB, COX-2 and MMP-9 expression increased significantly (P<0.05); Compared with the model rats, the AWR score of the Dajianzhong decoction group (60, 40 and 20 mmHg pressure) decreased significantly (P<0.05, P<0.01), ERK1/2, NF-κB, COX-2 and MMP-9 decreased significantly (P<0.05); Compared with the pinaverium bromide control group, ERK1/2, NF-κB, COX-2 and MMP-9 expression in the Dajianzhong decoction treatment group had no statistical difference (P>0.05).  Conclusion  Dajianzhong decoction can play a role in treating visceral pain of irritable bowel syndrome (IBS) by interfering ERK1/2/NF-κB pathway and its downstream molecules.
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    Monocarboxylate transporter 1 enhancing the expression of inducible nitric oxide synthase within M1 phenotype microglia under low-glucose condition 
    YU Zhe-cheng ZHANG Xiao-yan DU Juan-juan ZHOU Peng
    2022, 53 (3):  288-294.  doi: 10.16098/j.issn.0529-1356.2022.03.003
    Abstract ( )   PDF (4013KB) ( )  
    Objective To testify the link between the increase of inducible nitric oxide synthase (iNOS) and monocarboxylate transporter 1 (MCT1) expression in the M1 phenotype microglia under ischemic condition.   Methods  Photothrombosic ischemia stroke model was applied in 6 mice. BV2 cells were treated with low glucose medium contained 5mmol/L glucose for 2 hours. Immunofluorescent staining and Western blotting were used to check the responses from microglia in the mouse brains subjected to ischemia and BV2 cells were treated with low-glucose treatment. Application of RNA interference or plasmid transfection were used to regulate the MCT1 expression in BV2 cells.   Results  The expression levels of iNOS, MCT1 and arginase-1 (ARG1) increased in the ischemic side compared to the non-ischemic side of mice brain(P<0.01). In the BV2 cells exposed to low-glucose condition, iNOS and MCT1 levels increased(P<0.001), whereas ARG1 level decreased(P<0.01). RNA interference interfered the expression of MCT1 and then decreased the iNOS expression(P<0.01), while overexpression of MCT1 through plasmid transfection increased iNOS expression(P<0.01), while the ARG1 expressions in both conditions were not changed significantly.   Conclusion  After microglia polarized into inflammatory phenotype during ischemia period, iNOS production is related with MCT1 expression, and MCT1 is in the upstream of iNOS pathway.
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    Effects of dexmedetomidine on nuclear factor-κB inhibitor protein kinase/nuclear factor-κB inhibitor protein α/nuclear factor-κB pathway and cognitive dysfunction in rats with post-traumatic stress disorder
    CONG Hai-tao DING Jin-feng HE Hai-juan CHEN Ling-yang YANG Yan-qing ZHU Guang-qiu
    2022, 53 (3):  295-301.  doi: 10.16098/j.issn.0529-1356.2022.03.004
    Abstract ( )   PDF (3526KB) ( )  
    Objective To investigate the effects of dexmedetomidine (DEX) on the nuclear factor-κB inhibitor protein kinase (IKK)/nuclear factor-κB inhibitor protein α (IκBα)/nuclear factor-κB (NF-κB) pathway and cognitive dysfunction in rats with post-traumatic stress disorder (PTSD).   Methods Rats were randomly divided into control group, model group, positive group and DEX group. Except for the control group, the PTSD model was constructed by single prolonged stress method  (SPS), and the corresponding drugs were given after the completion the model. Open field test and Morris water maze method  were used to detect the autonomous activity, learning and memory ability of rats; HE staining was used to observe the morphological characteristics of cerebral cortex and hippocampus; ELISA and Western blotting were used to detect the contents of interleukin (IL)-1β, IL-6, tumor necrosis factor α(TNF-α) and the expression levels of IKK, IκBα, purinergic ligand-gated ion channel 7 receptor (P2X7R), leucine-rich repeat domain protein 3(NALP3) in hippocampus; the NF-κB activity was assessed by electrophoretic mobility shift assay (EMSA).   Results Compared with the control group, the cerebral cortex and hippocampal CA1 region of model group were in structural disorders, nuclear pyysis and other pathological changes happend, learning and memory ability of rats decreased(P<0.05), contents of IL-1β, IL-6 and TNF-α, expression levels of IKK, IκBα, P2X7R and NALP3, NF-κB activity in hippocampus increased(P<0.05); Compared with the model group, the pathological phenomena in cerebral cortex and hippocampal CA1 area of rats in positive group and DEX group were in alleviated, and the changes of the above indexes were opposite to those of model group(P<0.05).   Conclusion DEX can significantly improve the autonomous activity ability and learning and memory ability in PTSD rats, reduce inflammatory reaction in hippocampus and improve cognitive dysfunction, which may be related to the down-regulation of IKK/IκBα/NF-κB pathway.
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    A stereological observation of nuclear volumes and diameters of hippocampal neurons in aging rats
    GUO Zhen-zhou YANG Liang PENG Bin GUO Yang LI Yang-you HUANG Jing YANG Zheng-wei
    2022, 53 (3):  302-308.  doi: 10.16098/j.issn.0529-1356.2022.03.005
    Abstract ( )   PDF (1563KB) ( )  
    Objective To determine if there is atrophy of neuronal nuclei in the aging rat hippocampus.  Methods  The entire hippocampal formation in a cerebral hemisphere (on a random side) was removed from normal male SD rats aged 1, 2 and 3 years (each age-group 5 rats), serial sections (section thickness 40 μm) were cut after embedding in a resin of glycol methacrylate, and 3-4 sections were systematically sampled and stained with periodic acid, Schiff’s reagent and hematoxylin. The nuclear sizes of hippocampal neurons were estimated with stereological methods, first, the nuclei were sampled in a uniform random way using the optical disector; then, a Feret diameter of the largest nuclear profile at its focal plane (optical section) was measured as the nuclear diameter, and the nuclear volume was measured at the clearest focal plane of the nucleolus with the nucleator.   Results The mean diameters and volumes of the neuronal nuclei in the whole hippocampus were 8.7-9.2 μm and 392-457 μm3, respectively, in the 3 age-groups, without significant statistical difference between groups. There were no significant difference between groups, either, in the nuclear diameter or volume in each hippocampal region——the CA (hippocampus proper), dentate gyrus or subiculum.   Conclusion  In general, no age-related changes have been observed in the nuclear sizes of neurons in the whole hippocampus in the rat from middle to old age.
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    Cancer Biology
    Imiquimod inhibiting the proliferation of gliocytoma U87 cells by down-regulating STAT3/nuclear factor-κB signal pathway
    WANG Shu YIN Lu LIU Hong-bin XU Jia-zhi ZHAO Ji-bo PAN Yun-zhi SUN Yu-rong
    2022, 53 (3):  323-329.  doi: 10.16098/j.issn.0529-1356.2022.03.008
    Abstract ( )   PDF (3876KB) ( )  
    Objective  To explore the effect of imiquimod (IMQ) on the proliferation of  glioma U87 cell line U87.   Methods  U87 cells were divided into control group, 1 mmol/L I mmol/L MQ group, 5mmol/L IMQ group, 1 mmol/L IMQ+STAT3 inhibitor(STAT3-IN) group and 5 mmol/L IMQ+STAT3-IN group. To detect the number of 5-ethynyl-2’-deoxyuridine(EdU)-labeled cells or proliferation absorbance(A) values in each group by EdU and MTT assays. Interleukin(IL)-6 mRNA, tumor necrosis factor(TNF)-α mRNA and protein content in U87 cells of each group were detected by Real-time PCR or ELISA. Western blotting was used to detect the protein expression of STAT3, phosphorylated STAT3 (p-STAT3), nuclear factor(NF)-κB and phosphorylated NF-κB (p-NF-κB) in U87 cells of each group.  Results  Compared with the control group, the number of EdU-labeled cells and absorbance values of U87 cells were successively decreased in 1mmol/L IMQ group and 5mmol/L IMQ group, showing a dose-dependent manner (P<0.01, n=10). However, the number of EdU-labeled cells and the A values in IMQ+STAT3-IN group and 5 mmol/L IMQ+STAT3-IN group were significantly reduced. Compared with the control group, the protein expression of STAT3, p-STAT3, NF-κB, p-NF-κB, IL-6 and TNF-α were continuing low level in U87 cells of 1mmol/L IMQ group and 5mmol/L IMQ group (P<0.01, n=10). As well as in 1 mmol/L IMQ+STAT3-INgroup and 5 mmol/L IMQ+STAT3-IN group, the proteins of above were low expressed (P<0.01, n=10).   Conclusion Imiquimod decreased the contents of IL-6 and TNF-α by down-regulating STAT3/NF-κB pathway, and thus inhibited the proliferation of U87 cells.
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    Neurobiology
    Role of zinc finger protein 36, C3H type-like 1 mediating astrocytes activation in motor neuron degeneration in amyotrophic lateral sclerosis
    DING Kang ZHANG Feng-ping QI Gao-xiu LIN Meng CHEN Min CHEN Yan-chun GUO Zhang-yu ZHOU Feng-hua GUAN Ying-jun
    2022, 53 (3):  273-280.  doi: 10.16098/j.issn.0529-1356.2022.03.001
    Abstract ( )   PDF (5027KB) ( )  
    Objective To investigate the role of zinc finger protein 36,C3H type-like 1(ZFP36L1)mediating astrocytes activation in the degeneration of motor neurons in amyotrophic lateral sclerosis (ALS).  Methods Superoxide dismutase 1 (SOD1)-G93A transgenic mice were used as animal models, the wild-type littermates as the control (13 mice were taken from mutant and wild-type mice at each time point). The ZFP36L1 mRNA and protein levels of the spinal cord in the early, middle and late stage were detected by Real-time PCR and Western blotting. The expression and distribution of ZFP36L1 in the spinal cord were detected by immunofluorescence. Primary astrocyte model was established from 15 postnatal 1-2 day mice. The ZFP36L1 mRNA and protein levels in astrocytes were detected by Real-time PCR and Western blotting. Si-ZFP36L1 was transfected into SOD1-G93A mutant primary astrocytes. The transfection efficiency was detected by Western blotting. Tumor necrosis factor α (TNF-α) and interleukin-18 (IL-18) secreted from astrocytes after transfection were assessed by Western blotting and ELISA. After silencing ZFP36L1 in SOD1-G93A mutant primary astrocytes, it was cocultured with SOD1-G93A mutant NSC34 cells. 5’-ethyyl-2’deoxyuridine (EdU) test and the level of proliferating cell nuclear antigen (PCNA) were used to determine the effect of ZFP36L1 on NSC34 cell proliferation. TUNEL test and the level of cleaved-Caspase-3 were used to determine the effect of ZFP36L1 on NSC34 cell apoptosis. Blank small interfering RNA(siRNA) was transfected as the control group.   Results Compared with the wild-type mice, the mRNA and protein levels of ZFP36L1 were downregulated in the spinal cord of SOD1-G93A transgenic mice. In wild type mice, ZFP36L1 positive cells were mainly β-tubulin Ⅲ positive. In SOD1-G93A mutant mice, ZFP36L1 positive cells were mainly glial fibrillary acidic protein (GFAP) positive. The expression of ZFP36L1 in SOD1-G93A mutant primary astrocytes increased, and si-ZFP36L1 reduced the level of ZFP36L1 in SOD1-G93A mutant primary astrocytes significantly. Inflammatory factors including TNF-α, IL-18 decreased significantly after silencing ZFP36L1. In addition, after silencing ZFP36L1 expression, SOD1-G93A mutant primary astrocytes enhanced the proliferation activity of NSC34 cells and inhibited NSC34 cell apoptosis significantly.   Conclusion Astrocytes are activated in the process of ALS. ZFP36L1 promotes the degeneration of motor neurons in ALS through the inflammatory factors secreted by astrocytes.
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    Anatomy
    Superficial veins located in dorsolateral aspect of hemispheres on susceptibility-weighted imaging
    CHEN Zheng-zhen LI Chang-sheng XU Lin CHEN Dai-xi WEN Cai-yn CHEN Cheng-chun
    2022, 53 (3):  330-334.  doi: 10.16098/j.issn.0529-1356.2022.03.009
    Abstract ( )   PDF (1369KB) ( )  
    Objective To investigate whether physiological asymmetry of normal bilateral superficial veins located in dorsolateral aspects of hemispheres is existed on susceptibility-weighted imaging (SWI).   Methods  There were 45 healthy young volunteers in this study. After exclusion of intracranial lesions by using common brain magnetic resonance imaging examination, they underwent SWI examination. We observed the number, diameter and signal intensity of superficial veins located in dorsolateral aspects of hemispheres according to the images of SWI on transverse sections through the centrum semiovale, trunk of corpus callosum, splenium of corpus callosum, and superior colliculus.  Results  The frequencies of asymmetry of the bilateral superficial veins located in dorsolateral aspects of hemispheres on transverse sections through the centrum semiovale, trunk of corpus callosum, splenium of corpus callosum, and superior colliculus were 20.0%, 37.8%, 35.6% and 26.7%, respectively. The frequencies of mild asymmetry of the bilateral superficial veins located in dorsolateral aspects of hemispheres on above transverse sections were 17.8%, 31.1%, 31.1% and 24.4% respectively, and the frequencies of prominent asymmetry were 2.2%, 6.7%, 4.4% and 2.2%, respectively. According to chi square test, there was no gender difference in asymmetry of the bilateral superficial veins located in dorsolateral aspects of hemispheres.  Conclusion  SWI technique was used to show the physiological asymmetry of the bilateral superficial veins located in dorsolateral aspects of hemispheres on different transverse sections. Most of them are mild. Therefore, when cerebrovascular disease occurs, if transverse sections of images by SWI show asymmetry of the bilateral superficial veins located in dorsolateral aspects of hemispheres, the possibility of physiological phenomenon is needed to take into consideration.
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    Morphological analysis and measurement of spinoglenoid notch of scapula based on 3D reconstruction of CT
    LI Ming-bo CHEN Xiao-ying ZHENG Hai-xia
    2022, 53 (3):  335-339.  doi: 10.16098/j.issn.0529-1356.2022.03.010
    Abstract ( )   PDF (1772KB) ( )  
    Objective  To observe the anatomical structure of spinoglenoid notch of scapula by 3D reconstruction of normal adult scapula by computed tomography (CT), and to provide reference for clinical assessment of suprascapular nerve compression risk, suprascapular nerve compression treatment and shoulder joint operation.   Methods  Totally 335 cases of normal adult scapula were reconstructed by CT, and classified according to the anatomical shape of spinoglenoid notch; the spinoglenoid notch width (MN), spinoglenoid notch depth (OP), spinoglenoid notch thickness (XY), spinoglenoid notch angle (∠MON), distance from O point to the inner upper corner of scapula (OA), distance from O point to medial lateral edge of scapula (OB), distance from O point to inferior angle of scapula (OC) and distance from O point to the lowest point of suprascapular notch (OD) were observed and analyzed.   Results  1. The morphology of spinoglenoid notch was divided into four types: U type (41.79%), fin type (42.99%), L type (8.36%) and ladder type (6.86%). U type and fin type were the most common types. Comparison of the four shapes: fin type was the narrowest (11.58 ± 1.74) mm and the deepest (14.58 ± 1.81) mm, the ∠MON was the smallest (45.62 ± 6.43) ° and the ladder type was the widest (14.20 ± 2.67) mm and  the shallowest (10.80 ± 0.79) mm, the ∠MON was maximum (57.69 ± 2.22) ° and the least prone to suprascapular nerve compression. 2. There was no significant difference in MN, OP, XY, ∠MON, OA, OB, OC and OD between left and right sides. 3. The data of MN, OP, XY, OA, OB, OC and OD of men were larger than those of women significantly, but ∠MON was smaller than that of women, indicating that men’s spinoglenoid notch was thicker, wider and deeper, and scapula was wider and longer than that of women.    Conclusion  The measurement of the morphological and anatomical characteristics of spinoglenoid notch with CT three-dimensional reconstruction is helpful to evaluate the risk of suprascapular nerve compression, the treatment of suprascapular nerve compression, and provide guidance for clinical shoulder surgery.
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    Cancer Biology
    Effects of acetyl-CoA carboxylase 1 on proliferation, migration and invasion of glioma cell line U87
    WANG Yang ZHAO Bao-sheng LI Chao-hong LIU Yu-zhen
    2022, 53 (3):  317-322.  doi: 10.16098/j.issn.0529-1356.2022.03.007
    Abstract ( )   PDF (1782KB) ( )  
    Objective To explore the effect of acetyl-CoA carboxylase 1(ACC1) on cell proliferation, migration and invasion of human glioma cell line U87.   Methods  Western blotting was performed to examine endogenous ACC1 expression in human glioma cell lines U87, U251 and U373. ACC1 overexpression plasmid and the plasmid vector were transiently transfected into U87 cells. The level of ACC1 in control and ACC1 overexpression cells was examined by Western blotting. The effect of ACC1 on U87 cells migration and invasion was detected by Transwell assay. The effect of ACC1 on U87 cells scratch healing ability was detected by scratch test. The effect of ACC1 on U87 cells proliferation was investigated by MTT assay. Western blotting was conducted to detect the level changes of proteins.  Results  Among three human glioma cell lines U87, U251 and U373, endogenous ACC1 level in U87 cells was lower than that in other two cell lines. ACC1 overexpression inhibited U87 cell proliferation, as well as cell migration, invasion and scratch healing ability (P<0.05). Vimentin, fibronectin, urokinase type plasminogen activator (uPA), Bcl-2, cyclin B, cyclin D and p-STAT3 were down-regulated (P<0.05), P21 was up-regulated (P<0.05) after ACC1 overexpression.    Conclusion  These results suggest that ACC1 suppresses the proliferation, migration and invasion of human glioma cells, probably by inhibiting STAT3 activity.
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    Expression of mitogen-activated protein kinase kinase 1 in gastric cancer and its clinical significance
    ZHAO Yuan TAO Yue-jia YU Guang-fu GUO Jia-xin LI Meng-qi XU Yuan-yi HUANG Yun-ning
    2022, 53 (3):  309-316.  doi: 10.16098/j.issn.0529-1356.2022.03.006
    Abstract ( )   PDF (7742KB) ( )  
    Objective To investigate the expression of mitogen-activated protein kinase kinase 1 (MAP2K1) in gastric cancer and its clinical significance.   Methods  Immunohistochemistry and Western blotting were used to detect the protein expression of MAP2K1 in gastric cancertissues and cells. The morphology and the expression position of MAP2K1 were observed by immunofluorescence. MAP2K1 mRNA expression in gastric cancer tissues was analyzed by data mining of Starbase database and Oncomine database. The correlation between MAP2K1 mRNA expression and clinicopathological features was analyzed by UALCAN database. Survival analysis was performed using Kaplan Meier-Plotter online analysis tools. GEPIA2 database mining the relationship between MAP2K1 and gastric cancer stem cell related factors and drug resistance related factors.   Results  Immunohistochemistry, immunofluorescence and Western blotting showed that MAP2K1 protein was highly expressed in gastric cancer tissues and cells, and MAP2K1 was expressed in the cytoplasm of gastric cancer. According to the analysis of various databases, the expression of MAP2K1 mRNA in gastric cancer tissue was higher than that in normal gastric tissue, and the expression of MAP2K1 mRNA was closely related to gastric cancer stage, grade, lymph node metastasis and patient gender, and the overall survival rate of gastric cancer patients in the group with high MAP2K1 mRNA expression was significantly lower than that in the group with low MAP2K1 mRNA expression, which may be related to the characteristics of gastric cancer stem cells and drug resistance.   Conclusion  MAP2K1 is highly expressed in gastric cancer, and its expression level may affect the poor prognosis of patients by regulating stem cell related factors and drug resistance related factors. MAP2K1 may be a new diagnostic marker to determine the prognosis of gastric cancer patients.
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