Table of Content

2022, Volume 53 Issue 6
06 December 2022 Previous Issue    Next Issue
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Cell and Molecules Biology
Cancer Biology
Anthropology
Anatomy

Histology,Embryology and Developmental Biology


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Cell and Molecules Biology

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Regulation effect of endoplasmic reticulum stress and unfolded protein response activation on skeletal muscle fiber immune behavior HUANG Jing-wen ZHANG Ren-fei JIAN Xiao-ting LIAO Zhao-hong LAN Hai-qiang HUANG Tao HU Ji-jie LIAO Hua 2022, 53 (6):  727-736.  doi: 10.16098/j.issn.0529-1356.2022.06.006 Abstract ( )   PDF (11999KB) ( )   Objective To investigate the regulatory effects of activated endoplasmic reticulum stress (ERS) and unfolded protein response (UPR) on the immune behavior of the stressed muscle fibers in inflammatory environments induced by interferon-γ(IFN-γ).   Methods The myogenic precursor cells (MPCs) of C57 BL/6 mice cultured in vitro were differentiated into multinucleated myogenic tubes by horse serum and then to set up: 1. Control group; 2. IFN-γ group; 3. Tunicamycin group; 4. Thapsigargin group; 5. IFN-γ and 4-phenylbutyrate(4-PBA) combined treatment group; 6. IFN-γ, TG and 4-PBA combined treatment group; 7. IFN-γ and 4μ8c combined treatment group; 8. IFN-γ, TG and 4μ8c combined treatment group; 9. IFN-γ and GSK2606414 combined treatment group; 10. IFN-γ, TG and GSK2606414 combined treatment group. The level of myokines gene was detected by Real-time PCR. The expression of UPR key molecules including eukaryotic intiatio factor 2α(eIF2α), inositol requrring enzyme 1α(IRE1α) and activating transcription factor 6(ATF6) in muscle fibers was observed by immunofluorescence. Western blotting was used to detect immune molecules related to muscle cells, myokines and key molecules of UPR. Luminex analyzed the levels of pro-inflammatory myokines in muscle fibers.   Results The expression of H-2Kb, H2-Ea, Toll like receptor 3(TLR3), p-eIF2α and p-IRE1α were up-regulated in IFN-γ induced inflammatory environment. The expression of H-2Kb, H2-Ea, TLR3 and myokines in the group with UPR inhibitor 4-PBA was down-regulated compared with IFN-γ group, and the expression of these molecules in the group with IRE1α specific inhibitor 4μ8c was down-regulated compared with the IFN-γ group. The addition of protein kinase R-like endoplasmic eticulum(PERK) specific inhibitor GSK2606414 showed no significant change.  Conclusion In IFN-γ induced inflammatory environment, the UPR-IRE1α pathway activates and inhibits the synthesis of muscle fiber immune-related molecules, which further inhibits the muscle fiber mediated immune response and facilitates muscle regeneration. References | Related Articles | Metrics

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Relationship between the G894T polymorphism of endothelial nitric oxide synthase gene and lipid metabolism among hypertensive disorder complicating pregnancy patients LI Jing-yun LU Yun-hua ZHENG Li-xia ZHAO Gang LIU Hui-li 2022, 53 (6):  737-743.  doi: 10.16098/j.issn.0529-1356.2022.06.007 Abstract ( )   PDF (1125KB) ( )   Objective To investigate the relationship between the G894T polymorphism of the endothelial nitric oxide synthase (eNOS) gene and the lipid metabolism in patients with hypertensive disorder complicating pregnancy (HDCP).   Methods The 528 cases of HDCP patients admitted to the Xingtai Third Hospital from January 2016 to January 2020 were selected as the research objects, and 128 normal pregnant women during the same period were selected as the control group. The fasting peripheral venous blood of all study subjects in the early morning was collected, and blood lipid indexes, cystatin C (CysC) and uric acid levels and other biochemical index levels were measured. According to the blood lipid level, it was divided into normal blood lipid group and dyslipidemia group. The dyslipidemia group included 4 subgroups ［hyper triglyceride (TG) blood group (TG≥1.70mmol/L), low high-density lipoprotein cholesterol (HDL-C)］ hyper lipidemia group (HDL-C＜1.04mmol/L), high total cholesterol (TC) group (TC≥5.18mmol/L), mixed hyperlipidemia group (TG≥1.70mmol/L, TC≥ 5.18mmol/L)］. In addition, polymerase chain reaction-restriction endonuclease length fragment polymorphism (PCR-RFLP) was used to perform genotyping analysis on the G894T locus of    the eNOS gene, which was divided into TT, GT and GG genotypes. The distribution of gene polymorphisms in people with different blood lipid levels was compared, and Logistic regression was used to analyze the influencing factors of dyslipidemia and the relationship between NOS gene G894T polymorphism and dyslipidemia.   Results The expected and observed genotypes of GG, GT, TT, eNOS gene G894T locus in the study group and the control group conformed to the Hardy-Weinberg equilibrium law (χ2=0.651, P=0.722; χ2=1.845, P=0.398), and the GG type gene frequency and G allele frequency of the study group were higher than those of the control group, and the TT type gene frequency and T allele frequency of the study group were lower than those of the control group (P<0.05); homocysteine (Hcy), uric acid, C-reactive protein (CRP).The urine microalbumin to creatinine ratio (UACR) level and body mass index(BMI) were higher than those in the normal blood lipid group, and the rate of taking antihypertensive drugs was lower (P<0.05). Dyslipidemia group eNOS gene (G894T) gene GG gene frequency and G allele were higher than normal blood lipid group, TT gene frequency and T allele were lower than normal blood lipid group (P<0.001). The BMI of patients in the low HDL-C group, hyper TG group and mixed hyperlipidemia group was higher than that   of the normal blood lipid group (P<0.05); The Hcy level of the mixed hyperlipidemia group was higher than that of the normal blood lipid group (P<0.05). The TT gene frequency and T allele frequency of the low HDL-C group and the mixed hyperlipidemia group were significantly lower than those of the normal blood lipid group, and the GG gene frequency and G allele frequency were higher than the normal blood lipid group (P<0.05). Logistic regression analysis showed that genotype TT was a protective factor for dyslipidemia in patients with HDCP, and genotype GG, high BMI and high Hcy levels were independent risk factors for dyslipidemia.   Conclusion There is a significant correlation between the G894T polymorphism of the eNOS gene and dyslipidemia among patients with hypertension in pregnancy. The genotype TT is a protective factor, and the genotype GG/GT is an independent risk factor; at the same time, BMI and Hcy will also affect dyslipidemia. References | Related Articles | Metrics

Cancer Biology

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Expression and clinical significance of phosphoglycerate kinase 1 in hepatocellular carcinoma based on bioinformatics methods GUAN Cheng-jian YU Hua-jing ZHANG Xiao-dong RAO Quan ZHANG Wei-tao LIU Kun WU Hong-wei WANG Dong ZHANG Zhong-tao GUO Wei 2022, 53 (6):  744-753.  doi: 10.16098/j.issn.0529-1356.2022.06.008 Abstract ( )   PDF (11059KB) ( )   Objective To investigate the role of phosphoglycerate kinase 1 (PGK1) in tumorigenesis and its potential post-translational modification sites were investigated by bioinformatics method  and molecular biology experimental techniques, in order to provide evidence for PGK1 as a hepatocellular carcinoma(HCC) diagnostic biomarker and therapeutic target.   Methods From pan-cancer’s point of view, 10 967 samples were obtained from the cancer genome database TCGAs, and the expression of PGK1 in different tumors was explored by using cBioPortal and UALCAN analysis tools; Focusing on HCC, the expression differences of PGK1 in hepatocellular carcinoma tumor tissues and normal tissues were further analyzed by using GEO database analysis, Real-time PCR, Western blotting and cell invasion assay；The String database was used to analyze the protein-protein interaction network and gene set enrichment analysis; The CSS-Palm database and bioinformatics method  were used to predict protein post-translational modification sites on PGK1.   Results The PGK1 gene was abnormally amplified and overexpressed in various solid tumors, including hepatocellular carcinoma, and overexpression of PGK1 was correlated with a poor prognosis in hepatocellular carcinoma. Multiple novel post-translational modifications were existed on PGK1.   Conclusion PGK1 is closely related to the occurrence and development of various cancers including HCC and glycolytic metabolism abnormalities. Epigenetic modifications can regulate PGK1 and affect its cellular function in HCC. References | Related Articles | Metrics

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Effect of microRNA-221 targeting tissue inhibitor of metalloproteinase-2 to mediate Akt/mTOR signaling pathway on non-small cell lung cancer transplanted tumor mouse model LI Hui ZHANG Li-na SONG Rui-jia SONG Xiang-quan CHU Hui-ying ZHAO Li-yan 2022, 53 (6):  754-761.  doi: 10.16098/j.issn.0529-1356.2022.06.009 Abstract ( )   PDF (6212KB) ( )   Objective To explore the effects of miR-221 on tumor cell proliferation, migration and invasion in non-small cell lung cancer (NSCLC) xenograft model mice, and to preliminarily analyze its possible mechanism of regulating Akt/mammalian target of rapamycin(mTOR) signaling pathway by targeting tissue inhibitor of metalloproteinase-2 (TIMP-2) on tumor cells in non-small cell lung cancer (NSCLC) through tumor-bearing nude mice.   Methods The A549 cells were divided into control group, mimic group, TIMP-2 group and mimic+TIMP-2 group. The mimic group and TIMP-2 group were transfected with miR-221 mimic and TIMP-2 overexpression plasmids, respectively. The mimic+TIMP-2 group was simultaneously transfected with miR-221 mimic and TIMP-2 overexpression plasmids. The control group was transfected with the same amount of negative control plasmid. After transfection, the cells of each group were injected subcutaneously into the left forelimb to construct the corresponding 4 groups of NSCLC mouse models. The proliferation-related protein (Ki67) was detected by immunohistochemical staining to detected the effect of cell proliferation ability. Matrix metalloproteinase-2(MMP-2) and N-cadherin proteins in each group were tested by Western blotting to assess and compare the abilities of migration and invasion. The levels of miR-221, TIMP-2 and Akt/mTOR pathways in bone marrow and tumor tissues were detected by Real-time PCR and Western blotting.   Results When co-transfected with wild type(WT)-TIMP-2 and miR-221 mimic, the relative luciferase activity in the cells reduced significantly (P<0.05). Compared with the control group, the tumor mass, volume, Ki67, MMP-2 and N-cadherin protein expression levels, miR-221 and Akt/mTOR pathway levels were increased significantly, while the levels of TIMP-2 mRNA and protein were significantly reduced in the mimic group (P<0.05). Compared with the control group, the levels of TIMP-2 mRNA and protein in the TIMP-2 group increased significantly, while the other indicators decreased significantly (P<0.05). Tumor tissue mass, volume, Ki67, MMP-2, N-cadherin, miR-221 and Akt/mTOR pathway levels in mimic+TIMP-2 group were significantly lower than those in the mimic group and significantly higher than those in the TIMP-2 group, while TIMP-2 mRNA and protein levels were significantly higher than those in the mimic group and significantly lower than those in the TIMP-2 group (P<0.05).   ConclusionIn the NSCLC transplanted tumor mouse model, miR-221 may mediate the Akt/mTOR pathway by targeting the expression of TIMP-2 protein to promote cell proliferation, migration and invasion. References | Related Articles | Metrics

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MicroRNA-128-3p inhibiting epithelial-mesenchymal transition of ovarian cancer cells by regulating zince finger E-bonx binding homeobox 1 XU Yu-hong ZHANG Hui-ya WANG Yun-gen CHEN Jun-xia 2022, 53 (6):  762-768.  doi: 10.16098/j.issn.0529-1356.2022.06.010 Abstract ( )   PDF (8405KB) ( )   Objective To investigate the effect of mircoRNA-128-3p (miR-128-3p) on epithelial-mesenchymal transition (EMT) of ovarian cancer cells and its regulatory mechanism on zinc finger E-box binding homobox 1(ZEB1).   Methods Real-time PCR technology was used to detect the expression of miR-128-3p in epithelial ovarian cancer tissue (EOC) and adjacent normal tissue(30 cases each), and to observe whether there was a difference. Two human ovarian cancer cell lines, SKOV3 and A2780, were selected and transfected respectively. MiR-128-3p mimic (miR-128-3p mimic group) and negative control mimic (NC mimic group) were used to detect the expression of miR-128-3p in 4 groups by Real-time PCR to verify the interference effect. Transwell assay was used. The migration and invasion abilities of the four groups of cells were observed. The regulatory relationship between miR-128-3p and ZEB1 was verified by dual luciferase assay, and the expression level of ZEB1 protein after overexpression of miR-128-3p was detected by Western blotting; pcDNA-ZEB1 was transfected into SKOV3 and A2780 cell lines to make it overexpression of ZEB1 was divided into NC mimic group, miR-128-3p mimic group, and miR-128-3p mimic+pcDNA-ZEB1 group. Western blot  ting was used to detect the EMT-related protein E-cadherin in 6 groups of cells and the expression level of vimentin.   Results Real-time PCR result showed that the expression of miR-128-3p in EOC tissues decreased compared with that in adjacent tissues(P<0.01); The relative expression of miR-128-3p in the miR-128-3p mimic group was higher than that in the NC mimic group, while the numbers of migrating cells and invasive cells were lower than those in the NC mimic group (P<0.01). The result  of dual luciferase experiments showed that miR-128-3p had a negative regulatory effect on ZEB1. In SKOV3 and A2780 cells, the relative expression of ZEB1 protein in the miR-128-3p mimic group was lower than that in the NC mimic group, while the relative protein expression of Ecadhein was higher than that in the NC mimic group (P<0.01). The relative protein expression of E-cadhein in the miR-128-3p mimic+pcDNA-ZEB1 group was lower than that in the miR-128-3p mimic group (P<0.01). In SKOV3 and A2780 cells, the relative protein expression of vimentin in the miR-128-3p mimic group was lower than that in the NC mimic group, and the relative p  rotein expression of vimentin in the miR-128-3p mimic+pcDNA-ZEB1 group was higher than that in the miR-128-3p mimic group (P<0.01).   Conclusion The expression of miR-128-3p decreases in epithelial ovarian cancer tissues, which may be due to the regulation of ZEB1 to affect the expression of EMT-related proteins and participate in the EMT process of ovarian cancer cells. References | Related Articles | Metrics

Anthropology

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Facial changes of Mongolian nationality from youth to old age in China  LI Yong-lan ZHENG Lian-bin 2022, 53 (6):  793-801.  doi: 10.16098/j.issn.0529-1356.2022.06.014 Abstract ( )   PDF (857KB) ( )   Objective To explore the law of changes in the appearance of Chinese Mongolians with age.   Methods Totally 24 human indexes of head and face from 4410 cases (1916 males and 2494 females) and 13 Mongolian ethnic groups were measured, and 12 craniofacial indexes were calculated. Correlation analysis was used to study the changes of craniofacial indexes and indexes with age. The method  of analysis of variance was used to study whether the differences between the index values and index values of the three age groups were statistically significant.   Results With the increase of age, the distance between the two mandibular angles became wider, the mouth widened, the width, height and length of the nose increased, the skin of the upper lip increased, the length and width of the ear increased, the width of the head, forehead, face, mastoid and tragus narrowed, the width of the eyes narrowed, the upper and lower lips thinned, the circumference of the head and the height of the head decreased, and the length of the head did not change significantly. With the increase of age, the Mongolian face becomes narrower and longer, the proportion of the height of the face in the whole head high school increases, the nose becomes wider, and the head appears to be relatively longer, lower and wider. The distance between the left and right frontotemporal points is relatively closer, the mouth cleft appears more slender, and the ratio of length to width of the ear does not change much.   Conclusion With the increase of age, the facial features of Mongolian have changed. References | Related Articles | Metrics

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Types of tongue movements and their interrelationships in Kazakhs HUANG Ting LI Xin LAN Hui WENYou-feng 2022, 53 (6):  802-807.  doi: 10.16098/j.issn.0529-1356.2022.06.015 Abstract ( )   PDF (1541KB) ( )   Objective To investigate the types of tongue movement in Xinjiang Kazakhs, to explore the relationship between tongue movement types, and to provide inference for genetic of human anthropology.   Methods Totally 405 samples (178 males, 227 females) of Kazakh in Tacheng, Xinjiang, were investigated for the types of tongue movement. Chi-square test, u test and cluster analysis were used to explore the characteristics of tongue movement of Kazakhs and the tongue movement of other ethnic groups relation. Generalized multivariate dimension reduction (GMDR) analysis and logistic regression were used to analyze the correlations between tongue movements.   Results The frequencies of rolling tongue, folding tongue, twisting tongue, and pointed tongue were 73.08%, 41.97%, 18.02%, and 66.91% respectively. No gender difference was found in the frequencies of four tongue moving types. Compared with other ethnic groups in our country, the frequency of rolling tongue and pointed tongue was at a moderate level, the frequency of folding tongue was higher, and the frequency of twisting tongue was lower. The cluster analysis result  showed that the tongue movement type of Xinjiang Kazak was closest to that of Han nationality in Jinzhou, Liaoning. GMDR result  showed that among the four tongue movements, there were interactions between the combinations of rolling tongue and folding tongue, folding tongue and rolling tongue and pointed tongue, twisting tongue and pointed tongue, and rolling tongue and folding tongue，and pointed tongue and folding tongue. Logistic regression result  showed that there were correlations between rolling tongue and folding tongue, pointed tongue and folding tongue, folding tongue and pointed tongue and rolling tongue.  Conclusion Compared with the tongue movements of the domestic ethnic groups, the four types of tongue movements of the Kazakhs in Xinjiang are stronger in folding tongue movement, weaker in tongue turning movement, and average in rolling tongue and pointed tongue movement. There is a relationship between rolling tongue and folding tongue, pointed tongue and folding tongue, and folding tongue are related to pointed tongue and rolling tongue. References | Related Articles | Metrics

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Physical characteristics of Xibe adults  in Liaoning Province 2022, 53 (6):  808-814.  doi: 10.16098/j.issn.0529-1356.2022.06.016 Abstract ( )   PDF (946KB) ( )   Objective To study the physical characteristics of body of Xibe adults in Liaoning province and to accumulate data for the study of physical anthropology.   Methods The investigation method  and judging standard were refered to correlative regulations in “Anthropometric Method ”. The survey included 33 measurement items, 5 indicators and 6 types, 330 Xibe adult people (male 174, female 156) were recruited.  Results According to the classification of body part index, the rates of long trunk, wide chest, wide shoulder and wide pelvis were the highest in all the adults of Xibe nationality in Liaoning province, the rates of middle and sub-long legs were the highest in the males, and the rates of super short legs were the highest in the females.According to the stature type, the rates of high type were the highest in males and females.   Conclusion The body physical characteristics of male adults of Xibe nationality in Liaoning are the most similar to those of male adults of Xibe nationality in Xinjiang, and the body physical characteristics of female adults of Xibe nationality in Liaoning were between those of female adults of ethnic minorities in Northwest China and Northeast China. References | Related Articles | Metrics

Anatomy

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Anatomy of endoscopic odontoidectomy via submandibular approach WANG Yu-hui WANG Sheng-lin AN Feng-yang HU Wei-ni LUO Yan-lin ZHU Li MA Fu-rong 2022, 53 (6):  769-775.  doi: 10.16098/j.issn.0529-1356.2022.06.011 Abstract ( )   PDF (8134KB) ( )   Objective To provide the basis for endoscopic craniocervical junction surgery through cervical CT image and endoscopic odontoid process anatomy of atlas, axis and odontoid.   Methods A total of 150 cases of cervical vertebrae were selected for high resolution thin slice plain CT measurement to evaluate the atlantoaxial structure and its adjacent structure, and to estimate the safe boundary of odontoid process resection. The atlantoaxial odontoid process was anatomized on 3 cadaver head specimens under endoscope through the submandibular approach using STORZ endoscopy system and endoscopic surgical instruments.  Results The average length of atlas anterior arch and other anatomical marks were measured by CT, and the safety boundary area of odontoidectomy was estimated to be (240.9 ± 39.92)mm2, male: (248.3 ± 49.64)mm2, Female: (233.2 ± 24.54)mm2. Through the submandibular endoscopic approach, the atlantoaxial anatomy and odontoidectomy anatomy made a transverse incision at the midpoint of the connecting line between one mandibular angle and hyoid bone to expose the submandibular triangle area. Under the endoscope, the digastric muscle and the greater angle of hyoid bone were exposed through the submandibular triangle area, and the retropharyngeal space was passively separated layer by layer to the prevertebral space to expose the prevertebral fascia. After removing the prevertebral tissue, the atlas, the dentate process of the axis, the atlantooccipital joint, the atlantoaxial joint, and part of the foramen magnum were fully exposed.  Conclusion Estimating odontoid resection safety boundary area by CT image, in combination with endoscopic odontoidectomy anatomy via sunbmandibular approach, we can perform the surgery safely and efficiently under the bright of endoscope with surgical instruments, which can significantly reduce the incidence of cerebrospinal fluid leakage and postoperative infection while decompressing. References | Related Articles | Metrics

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Finite element analysis of facet joint between normal and lumbar disc herniation in adolescents WANG Yi-dan XU Yang-yang WANG Hai-yan LI Xiao-he LI Zhi-jun JIN Feng WU Chao HE Yu-jie GAO Ming-jie GUAN Huan-huan 2022, 53 (6):  776-784.  doi: 10.16098/j.issn.0529-1356.2022.06.012 Abstract ( )   PDF (1880KB) ( )   Objective To investigate the effect of facet joint asymmetry on lumbar biomechanics in normal and patients with adolescent lumbar disc herniation (ALDH).   Methods Mimics 21.0, 3-Matic Medical 13.0, Geomagic Wrap 2017, HyperMesh 2019 and finite element software ABAQUS 2021 were combined to establish three-dimensional finite element models of normal lower lumbar spine and adolescent lumbar disc herniation. According to the difference between the left and right facet joints, three cases of normal and ALDH  patients aged 13-18 years old were selected, with a total of 6 cases. The stress of intervertebral disc under different torque loads (neutral position, lateral flexion and rotation) was analyzed and compared.   Results 1. Three dimensional finite element models of L3-L5 segments were established in 6 male normal and adolescent patients with lumbar disc herniation, and the stress and displacement nephogram of lumbar disc herniation in normal neutral position, flexion, extension, lateral flexion and rotation were obtained; 2. The stress of L4-5 annulus fibrosus increased when the facet joint angle of normal adolescents was symmetrical in the neutral position; 3. The stress of annulus fibrosus was greater than that of nucleus pulposus under different conditions, and the stress of annulus fibrosus was flexion > neutral position > extension; 4. In patients with ALDH, the left side of the facet joint was larger than the right 10 ° model, and the stress in the posterior side of the annulus fibrosus of L4-5 segment increased significantly under extension condition. Under lateral flexion condition, the left stress of the left annulus fibrosus was compared with the right stress of the right annulus f  ibrosus, t=1.575, P<0.05, the difference was statistically significant, the right stress of the right annulus fibrosus was greater than the left stress of the left annulus fibrosus. Under the rotating condition, the stress on the left and right sides of the fiber ring was greater than that on the back side.   Conclusion 1. Compared with normal and patients with ALDH, the stress of intervertebral disc increases under different postures, which may increase the shear load of intervertebral disc and aggravate the process of intervertebral disc degeneration; 2. When the degree of left-right asymmetry of facet joint is more than 10 degrees, the stress on the side with small degree is greater. Facet joint asymmetry can lead to the overload of vertebral body and intervertebral disc, leading to spinal instability; 3. The stress of intervertebral disc in extension is less than that in neutral position and flexion. Extension can relieve the pressure of intervertebral disc and play an auxiliary role in the recovery of patients with ALDH. References | Related Articles | Metrics

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Expression of protein tyrosine kinase 7 and receptor tyrosine kinase-like orphan receptor 2 in the brainstem of amyotrophic lateral sclerosis transgenic mice with hSOD1-G93A mutation MENG Fan-di GUAN Ying-jun ZHAO Zhen-han CHEN Yan-chun LIU Jin-meng WANG Xue-mei ZHANG Hao-yun ZHOU Feng-hua 2022, 53 (6):  689-697.  doi: 10.16098/j.issn.0529-1356.2022.06.001 Abstract ( )   PDF (15965KB) ( )   Objective To investigate the relationship between changes in protein tyrosine kinase 7 (PTK7) and receptor tyrosine kinase-like orphan receptor 2 (ROR2) expression in the brainstem and the pathogenesis of amyotrophic lateral sclerosis (ALS).   Methods Forty-four human superoxide dismutase 1(hSOD1)-G93A mutant ALS transgenic mice were selected, and an equal number of wild-type littermates was used as control. The brainstems were isolated at day 70, day 95, day 108 and day 122 after birth, and the morphology of hypoglossal nucleus (12N) and nucleus of facial nerve (7N) neurons in the brainstem of the model mice were observed by Nissl staining. The mRNA and protein expression of PTK7 and ROR2 were detected by RT-PCR and Western blotting respectively, and the cellular localization and distribution of PTK7 and ROR2 in 12N and 7N were observed by immunofluorescence double-labeling technique.   Results The result  of Nissl staining showed that Nissl bodies in the neurons reduced distinctly with vacuolar degeneration of neurons, cell body atrophy and nuclear volume reduction in the 12N and 7N brainstems of ALS transgenic mice. RT-PCR result  indicated that ROR2 and PTK7 mRNA level in the brainstem of ALS transgenic mice were up-regulated at day 70, day 95, day 108 and day 122 compared with wild-type littermates. Western blotting result  showed that PTK7 protein was up-regulated at day 70,  day 95, day 108 and day 122, ROR2 protein was up-regulated at day 70, day 95, day 108, and down-regulated at day 122 in the brainstem of ALS transgenic mice compared with wild-type littermates. Immunofluorescence result  showed that ROR2/neuronal nuclei (NeuN)and PTK7/NeuN double positive cells, ROR2/glial fibrillary acidic protein (GFAP) and PTK7/GFAP double positive cells were observed in the 12N and 7N of the brainstem of ALS transgenic mice and wild-type mice, suggesting that ROR2 an  PTK7 were expressed both in neurons and astrocytes.   Conclusion PTK7 and ROR2 are abnormally expressed in the brainstem of ALS transgenic mice, which is closely related to the pathogenesis of ALS. References | Related Articles | Metrics

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Expression of β-site amyloid precursor protein cleaving enzyme 1 in rat superior cervical ganglion tissues FAN Ya-nan LI Chao-hong JIA Xiang-lei JIN Jia-hao LIU Yu-zhen 2022, 53 (6):  698-704.  doi: 10.16098/j.issn.0529-1356.2022.06.002 Abstract ( )   PDF (7866KB) ( )   Objective To observe the expression and localization of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in rat superior cervical ganglion and the effect of chronic intermittent hypoxia (CIH) on BACE1 level.   Methods The expression and distribution of BACE1 in superior cervical ganglion were detected by RT-PCR, Western blotting and immunohistochemistry.  Totally 16 male SD rats were randomly divided into control group and CIH group, 8 rats in each group. After 2 weeks of modeling, the effect of CIH on BACE1 and peroxisome proliferators activated receptor gamma co-activator 1 alpha (PGC-1α) mRNA level was detected by RT-PCR.    Results BACE1 was expressed in rat superior cervical ganglion, and mainly distributed in satellite glial cells and nerve fibers, but not in blood vessels, neurons and small intesely fluorecent(SIF) cells. CIH down-regulated BACE1 mRNA level, but up-regulated PGC-1α mRNA level (P<0.01).   Conclusion BACE1 is located in satellite glial cells and nerve fibers in the superior cervical ganglion of rats. The decreased level of BACE1 may be involved in the regulation of CIH-induced synaptic plasticity of superior cervical ganglion. References | Related Articles | Metrics

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Effect of the aqueous extracts of ganoderma leucocontextum on cognitive function of aging rats  WANG Yu QIN Xu HE Jiu-jun WANG Yi-xuan 2022, 53 (6):  711-718.  doi: 10.16098/j.issn.0529-1356.2022.06.004 Abstract ( )   PDF (11922KB) ( )   Objective To investigate the effects of the aqueous extracts of ganoderma leucocontextum (GLAE) on cognitive decline of aging rats and possible regulation mechanism.    Methods Fifty rats were divided into five groups, control group, model group, GLAE low-dose group, GLAE middle-dose group and GLAE high-dose group. Aging SD rat models were made by D-galactose, and then treated continuously with different doses (0, 50, 100, 200 mg/kg) of GLAE. The novel object recognition and step down test were performed to detect the changes of rats cognitive function. The brain tissue was stained with toluidine blue, Giemsa and HE staining and observed. The cerebral cell DNA damage was detected by comet assay. Expressions of protein kinase A (PKA)/cyclic adenosine monophosphate (cAMP)-response element binding protein (CREB) signaling pathway related factors in brain were respectively detected by ELISA, Western blotting and Real-time PCR.    Results Compared with the model group, administration of GLAE could obviously alleviate rats cognitive decline and pathological change. The levels of cell DNA damage reduced markedly (P<0.05). The contents of cAMP, brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF), and also expression levels of mRNA and protein of PKA, BDNF, NGF, CREB in the brain increased significantly in each medicated group (P<0.01, P<0.05).   Conclusion GLAE can improve cognitive function, and its mechanism may be related to activation of brain PKA/CREB signaling pathway, increase in neurotrophic factor content and inhibition of cell DNA damage.  References | Related Articles | Metrics

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Based on PI3K/Akt signaling pathway to explore the effects of olanzapine on cognitive function and neuronal damage in schizophrenic rats ZHONG Hai-long JIANG Xun-ying HE Zhu-fa MAO Zhong-zhen 2022, 53 (6):  719-726.  doi: 10.16098/j.issn.0529-1356.2022.06.005 Abstract ( )   PDF (2999KB) ( )   Objective To explore olanzapine  effect on the cognitive function and neuronal damage of aged schizophrenic rats based on the PI3K/Akt signaling pathway.   Methods Ten-week-old SD rats were randomly divided into a blank control group (n=12) and a modeling intervention group (n=48). The modeling group were injected with didroxapine maleate ［MK-801，0.2 mg/(kg·d)］ for 14 days. And the model was evaluated by general behavioral studies to determine the success of model building. The model rats were randomly divided into model group and low, medium, and high dose olanzapine groups ［10, 20, 40 mg/(kg·d)］, each with 12 rats. The control group and model group were given distilled water; the low, medium, and high dose olanzapine groups were given olanzapine for 21 days. The stereotyped lines were scored by the standard of Sams Dodd and Hoffman, the cognitive evaluation of the rats was performed by the Morris water maze, and the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the serum were determined by ELISA. The activities of dihydrokaempferol (Ach) and acetyl cholinesterase (AchE )in brain tissue were detected by acetylcholinesterase activity  assay kit. Rat brain tissue PI3K, Akt, mammalian target of rapamycin (mTOR) mRNA expression levels were detected by Real-time PCR.   Results Compared with the model group, the stereotyped behavior and ataxia scores, escape latency, number of crossing platforms, serum levels of IL-6, TNF-α, AchE, phosphorylated PI3K (p-PI3K), phosphorylated Akt (p-Akt) protein expression decreased (P<0.05 or P<0.01), while brain tissue Ach, PI3K, mTOR and phosphorylated mTOR (p-mTOR) protein content increased (P<0.05 or P<0.01 ) in the low, medium and high dose olanzapine groups. The content of Akt was increased in the low-dose group. Compared with the model group, Akt and mTOR mRNA in the brain tissue of rats in the low, medium, and high-dose alanzapine groups expression levels were down-regulated (P<0.05 or P<0.01). PI3K mRNA in the brain tissue of rats in the low, medium, and high-dose alanzapine groups expression levels were down-regulated (P<0.05 or P<0.01).  Conclusion Olanzapine can reduce stereotyped behavior and ataxia scores, escape latency, number of crossing platforms, IL-6, TNF-α, AchE and increase Ach content and regulate the PI3K/Akt signaling pathway to relieve the schizophrenia. References | Related Articles | Metrics

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Effect of electroacupuncture on the formation of glial scars in cerebral cortex of rats with focal cerebral ischemia/reperfusion XU Hong-bei LUO Yong 2022, 53 (6):  705-710.  doi: 10.16098/j.issn.0529-1356.2022.06.003 Abstract ( )   PDF (4863KB) ( )   Objective To observe the effect of electroacupuncture on the formation of glial scars in the cerebral cortex of rats with focal cerebral ischemia/reperfusion（I/R）.   Methods The focal cerebral ischemia/reperfusion model was established by thread method, and “Baihui” (GV 20) and left “Siguan” (Hegu LI 4/Taichong LR 3) were selected as acupoints points. Seventy-eight male SD rats were randomly divided into sham group (sham), focal cerebral ischemia/reperfusion (I/R), focal cerebral ischemia/reperfusion+ electroacupuncture group (I/R+EA). Seven days after reperfusion, modified neurological severity score (mNSS) was used to evaluate neurological deficit, HE staining was used to observe the degree of ischemic cerebral cortex. Immunofluorescencte was used to observe glial fibrillary acidic protein (GFAP)/neurocan, GFAP/phosphacan positive cells in the ischemic cerebral cortex. The average immunofluorescent intensity of GFAP, neurocan and phosphacan, and the levels of GFAP, neurocan and phosphacan mRNA in the ischemic cerebral cortex were analyzed by Real-time PCR, respectively.   Results Compared with the I/R group, mNSS score in I/R+EA group decreased significantly (P<0.01), and the degree of brain tissue damage reduced obviously. There were very few GFAP+/neurocan+ cells and GFAP+/phosphacan+ cells in the sham group. The astrocytes in I/R group were hypertrophy, increasing and thickened protrusions, more GFAP+/neurocan+ cells and GFAP+/phosphacan+ cells were dectected as well. While the count of GFAP+/neurocan+ cells and GFAP+/phosphacan+ cells in I/R+EA group were significantly lower than those in I/R group. In addition, the average immunofluorescence intensity of GFAP, neurocan and phosphacan in I/R+EA group, and mRNA content of above mentioned indicators were significantly lower than those in I/R group (P<0.05).   Conclusion Electroacupuncture promotes the recovery of neurological function in rats with focal cerebral I/R and reduce brain tissue damage, which may be related to the inhibition of glial scar formation in cerebral cortex.  References | Related Articles | Metrics

Histology,Embryology and Developmental Biology

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Effect of formyl peptide receptor 2 in recurrent spontaneous abortion through p38 MAPK pathway LI An-na FANG Zhen-ya ZHOU Mei-juan LI Shu-xian ZHAO Man GUO Jun-jun ZHANG Mei-hua 2022, 53 (6):  785-792.  doi: 10.16098/j.issn.0529-1356.2022.06.013 Abstract ( )   PDF (18444KB) ( )   Objective To explore the express of formyl peptide receptor 2(FPR2) in villi of recurrent spontaneous abortion (RSA), the effect on proliferation, migration and invasion of trophoblast, and the mechanism to clarify the effect of FPR2 on trophoblast function and explore its role and mechanism in recurrent spontaneous abortion.   Methods Clinical villus specimens of 30 normal and 30 RSA patients were collected. Immunohistochemical staining, Real-time PCR and Western blotting were used to detect the location and expression of FPR2 in villi of patients with RSA and normal pregnant women. CRISPR/Cas-9 technique was used to knock down FPR2 in HTR-8/SVneo cells, CCK-8 assay, wound healing and Transwell assays were used to determine the ability of cell viability, migration and invasion. Immunofluorescent staining and Western blotting were used to analyze the changes of phosphorylated p38 MAPK(p-p38 MAPK)/p38 MAPK protein expression after applying with p38 MAPK inhibitor SB203580 alone or in combination.   Results The expression of FPR2 in villi of patients with RSA increased. FPR2 knock-down improved the biological functions of HTR-8/Svneo cells such as proliferation, migration and invasion significantly. The expression of p-p38 MAPK was up-regulated significantly by FPR2 knock-down, and the ability enhancement of migration and invasion of trophoblasts was reversed partially by SB203580 which inhibits p38 MAPK pathway. FPR2 knock-down caused the change of p38 MAPK signaling pathway related to proteins.   Conclusion FPR2 is highly expressed in trophoblasts of RSA patients, and inhibits the migration and invasion of trophoblasts through p38 MAPK signaling pathway, which may play an important role in RSA.  References | Related Articles | Metrics