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2022, Volume 53 Issue 4
06 August 2022 Previous Issue    Next Issue
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Cell and Molecules Biology
Anatomy

Cell and Molecules Biology

Cancer Biology
Anthropology


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Cell and Molecules Biology

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Effect of exosomes derived from bone marrow mesenchymal stem cells on the polarization of mouse liver Kupffer cells QIN Yang-yang XU Long-fei WANG Qi HAN Jing HONG Yan 2022, 53 (4):  447-452.  doi: 10.16098/j.issn.0529-1356.2022.04.007 Abstract ( )   PDF (14264KB) ( )   Objective  To investigate the  the effect of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) on the polarization of liver Kupffer cells.     Methods  After BMSCs were isolated and cultured in vitro, their surface molecular expression was identified by flow cytometry, and their differentiation potential was induced and identified by osteogenic and lipogenic induction media. Exosomes were extracted from the supernatant of BMSCs culture by exosomes extraction kit, and their morphology was observed by electron microscopy, and surface molecular expression was identified by flow cytometry. In vitro cultured Kupffer cells were randomly divided into normal culture, lipopolysaccharide(LPS) stimulation and LPS co-culture groups, and morphological changes in Kupffer cells were observed under light microscope.Sixty mice were injected intraperitoneally with CCl4 to replicate acute liver injury model and randomized into PBS control and exosomes treatment groups, and the liver histopathology and the expression of liver functional alanine transferase(ALT) and aspartate aminotransferase(AST) were detected by HE staining. Western blotting was used to detect the  expressions of inducible nitric oxide synthase(iNOS) and arginase-1(ARG1) in the three groups of Kupffer cells cultured in vitro and the two groups of liver tissues in vivo. The expression of iNOS, ARG1, interleukin(IL)-1β, tumor necrosis factor(TNF)-α and C-X-C motif chemokine(CXCL)-10 were detected by Real-time PCR.    Results  BMSCs were successfully isolated and had the ability of differentiation of osteoblasts and adipocytes, and expressed CD105 and CD45. The exosomes were bubble-like and expressed CD63 and CD81. Light microscopy showed that exosomes of BMSCs attenuated Kupffer cells activation, and exosomes of BMSCs attenuated pathological changes of liver tissue after injection (P<0.05), decreased the high expression of ALT and AST in liver function (P<0.05); Western blotting showed that ARG1 expression increased in LPS and exosomes co-cultured in vitro and exosomes treated in vivo (P<0.05), iNOS decreased (P<0.05); Real-time PCR showed decreased expression of iNOS, IL-1β, TNF-α and CXCL-10 in LPS and exosomes co-cultured group and exosomes treated group (P<0.05), ARG1 expression increased (P<0.05).    Conclusion  Exosomes derived from BMSCs inhibited M1 type polarization of mouse Kupffer cells. References | Related Articles | Metrics

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Wnt7a/β-catenin/autoimmune regulator signaling pathway involved in the occurrence of type 1 diabetes LI Yuan-di MA Jing-yi YU Jia-shan HE Ke-ke WEN Ting-hao GAO Jie LI Hong SU Min HU Rong 2022, 53 (4):  453-460.  doi: 10.16098/j.issn.0529-1356.2022 Abstract ( )   PDF (9248KB) ( )   Objective  To investigate the relationship between the expression of Wnt signaling pathway, autoimmune regulator (AIRE) and type 1 diabetes (T1D) tissue specific antigen (TSAs) insulin 2(Ins2) and glutamic acid decarboxybase(GAD67) in thymus and the occurrence of T1D in NOD/Ltj mice with spontaneous type 1 diabetes (T1D).   Methods  Sixty female NOD/Ltj mice were divided into three groups: 3 weeks group, 16 weeks non-onset group and 16 weeks onset group. Two consecutive non-fasting blood glucose levels ≥ 11.1 mmol/L were considered as the occurrence of T1D. Pancreatic HE staining was used to observe the occurrence of islet inflammation. Anti-Ins and CD45 immunohistochemical staining showed islet β cells or infiltrating inflammatory cells. The protein levels and mRNA expressions of Wnt7a, β-catenin, AIRE, Ins and GAD67 in thymus were detected by Western blotting and Real-time PCR. The proportion of T cells in thymus was analyzed by flow cytometry.    Results  1. With the occurrence of T1D, the islet structure was destroyed, a large number of lymphocytes infiltrated, and the remaining islet cells were reduced. A large number of CD45+ cells were observed around Ins+ islet β cells. 2. The protein levels and mRNA expressions of Wnt7a, β-catenin, AIRE, Ins2 and GAD67 in thymus decreased with age. The expression of Ins in thymus decreased in the same week-old group compared with that in the control group. 3. Compared with the 3 weeks group, the proportion of CD4 and CD8 single positive T cells in the 16 weeks group was lower; In the 16 weeks group, the proportion of CD4 and CD8 single positive T cells increased, while the proportion of CD4 and CD8 double positive T cells decreased.   Conclusion  Wnt7a/β-catenin signaling pathway may be involved in the occurrence and development of T1D by regulating AIRE expression and down-regulating T1D-related TSAs expression. References | Related Articles | Metrics

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Effect of Wnt5b overexpression on the differentiation of embryonic liver stem cells in mice and its repair effect on chronic liver injury in mice JIANG Hao XIE Zhuo-jun ZHANG Hao-dong ZHOU Hong LUO Qing KANG Quan 2022, 53 (4):  461-469.  doi: 10.16098/j.issn.0529-1356.2022.04.009 Abstract ( )   PDF (22593KB) ( )   Objective  To investigate the effect of wingless-type MMTV integration site family member 5b(Wnt5b) gene overexpression mediated by recombinant adenovirus on the differentiation of mouse embryonic liver stem cells and repair of chronic liver injury in mice.    Methods  Recombinant adenoviruses expressing Wnt5b and green fluorescent protein (GFP) were applied respectively to infect mouse fetal liver stem cells HP14-19, and induced its differentiation and verified the expression of Wnt5b through Real-time PCR and Western blotting. It also applied indocyanine grean(ICG) uptake experiment and periodic acid-schiff(PAS) staining to detect the differentiation ability of HP14-19 into hepatocyte-like cells. The Real-time PCR was chosen to detect hepatocyte markers albumin (Alb) and cytokeratin 18 (CK18) expression. Forty-eight experimental male BALB/c mice were randomly divided into control group, model group, stem cell treatment group and Wnt5b modified stem cell treatment group. The carbon tetrachloride(CCl4) was selected to establish a chronic liver injury model. After 2 weeks of stem cell treatment, liver pathological staining and immunohistochemistry were used to evaluate liver repair.  Results  Both Real-time PCR and Western blotting detection showed that Wnt5b was successfully overexpressed in HP14-19（P<0.001）. The mRNA expressions of Alb and CK18 in the Wnt5b group were higher than those in the GFP group and control group by Real-time PCR（P<0.001）. ICG uptake test and PAS staining showed that the ICG uptake capacity and glycogen synthesis capacity of Wnt5b group increased significantly compared with the GFP group after 7 days of induction（P<0.01）. Liver HE staining result  showed that the model group had an obvious liver injury and liver tissue structure disordered. The stem cell group liver tissue structure partially recovered, and the liver tissue recovery in the Wnt5b group was better than that in the stem cell group. Masson staining showed that the model group had obvious fiber hyperplasia, and the Wnt5b group had less blue-stained fiber than the stem cell group（P<0.05）. The immunohistochemistry showed collagen type Ⅰ(ColⅠ) and myeloperoxidase (MPO) increased significantly in the  model group, and the repair effects of these indexes were significant in the Wnt5b group than in the fetal stem cell group（P<0.05）.    Conclusion  Wnt5b can induce mouse fetal liver stem cells to differentiate into mature hepatocyte-like cells. Also, the genetically modified fetal liver stem cells have more advantages in repairing the liver of chronic liver injury mice than fetal liver stem cells. References | Related Articles | Metrics

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Effects of levosimendan on the hypoxic/reoxygenated cardiomyocyte fibrosis by regulating long chain noncoding RNA/microRNA-641 HUANG Hong-ke LUO Jian-wei RAN Hua 2022, 53 (4):  479-487.  doi: 10.16098/j.issn.0529-1356.2022.04.011 Abstract ( )   PDF (8294KB) ( )   Objective  To investigate whether levosimendan (Lev) affects hypoxia/reoxygenation (H/R)- induced cardiomyocyte proliferation, apoptosis and fibrosis by regulating the molecular axis of long chain noncoding RNA (LncRNA) eosinophil granule ontogeny transcript(EGOT)/microRNA(miR)-641.    Methods  Rat cardiomyocytes H9C2 were cultured in vitro, and H/R-treated cells were used to establish cell damage models, which were randomly divided into control group, H/R group, H/R+Lev 1 μmol/L (H/R + Lev-L) group, H/R+Lev 5 μmol/L (H/R + Lev-M) group, and H/R+Lev 10 μmol/L (H/R + Lev-H) group,9 samples per group. MTT method was used to detect cell proliferation. Flow cytometry was used to detect the apoptosis rate. Real-time PCR was used to detect the expression levels of EGOT and miR-641 mRNA. PcDNA-EGOT and EGOT small interfering RNA(si-EGOT) were transfected into H9C2 cells respectively, and the cell proliferation and apoptosis rates were detected by the above method. The dual luciferase report experiment verified the targeting relationship between EGOT and miR-641. Western blotting was used to detect the expression levels of Bax, Bcl-2, collagen Ⅰ(ColⅠ), collagen Ⅲ(ColⅢ), tissue inhibitor of matrix metalloproteinase 2(TIMP2), matrix metalloproteinase-2(MMP-2).    Results   Compared with the control group, the cell survival rate of the H/R group reduced significantly (P<0.05), the apoptosis rate increased significantly (P<0.05), and the protein levels of Bax, ColⅠ, Col Ⅲ, TIMP2, and MMP-2 increased significantly (P<0.05), the level of Bcl-2 protein reduced significantly (P<0.05), the expression level of EGOT reduced significantly (P<0.05), the expression level of miR-641 increased significantly (P<0.05). Compared with the H/R group, the cell survival rate of the H/R + Lev-L group, H/R + Lev-M group, and H/R + Lev-H group increased significantly (P<0.05), and the apoptosis rate decreased significant (P<0.05),the protein levels of Bax, ColⅠ, ColⅢ, TIMP2, MMP-2 reduced significantly (P<0.05), the level of Bcl-2 protein increased significantly (P<0.05), the expression level of EGOT increased significantly (P<0.05), the expression level of miR-641 reduced significantly (P<0.05), and each index of H/R + Lev-L group, H/R + Lev-M group, H/R + Lev-H group, the difference was   statistically significant (P<0.05). The dual luciferase report experiment confirmed that EGOT ccould target and bind to miR-641. The effect of transfecting pcDNA-EGOT and Lev was similar. Transfection of si-EGOT could reduce the effect of Lev on H/R-induced proliferation, apoptosis and fibrosis of H9C2 cells.  Conclusion  Levosimendan may promote H/R-induced H9C2 cell proliferation and inhibit apoptosis and fibrosis by up-regulating EGOT expression and down-regulating miR-641 expression. References | Related Articles | Metrics

Anatomy

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Anatomic classification of the sustentaculum tali and its characteristics analysis ZHANG Lei LI Zhi-qing TANG Xiao-gao ZHOU Xin LI Bing-kun FU Shi-jie 2022, 53 (4):  515-519.  doi: 10.16098/j.issn.0529-1356.2022.04.015 Abstract ( )   PDF (2530KB) ( )   Objective  To explore the anatomical morphology and characteristics of the sustentaculum tali based on CT three-dimensional reconstruction, so to provide anatomical support for the treatment of calcaneal fractures with screw implantation.    Methods  From March 2019 to March 2020, a total of 336 adult calcaneal CT three-dimensional reconstruction images from the Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University were collected after exclusion of inclusion criteria. The CT threedimensional reconstruction of calcaneus was classified according to the β, the prolate axial intersection between sustentaculum tali and calcaneus on the normal posterior. Reference points, β; AB, the distance from the inside of sustentaculum tali to the inside of posterior talar articular surface; AC, the distance from the inside of sustentaculum tali to the outside of posterior talar articular surface; AD, the distance from the inside of sustentaculum tali to lateral of calcaneal; AE, the distance from the inside of sustentaculum tali to medial process of calcaneal tuberosity; AF, the distance from the inside of sustentaculum tali to calcaneal tuberosity; AG, the distance from the inside of sustentaculum tali to lateral process of calconeal tuberosity. The results were statistically analyzed according to type, sex and body side.    Results  Under the β, the sustentaculum tali was classified into three types: typeⅠ(β<70°, 68 cases, 20.24%), type Ⅱ(70°≤β<80°, 153 cases, 45.54%), type Ⅲ(80°≤β<90°, 115 cases, 34.23%). For the distance of β, AB, AF, there were statistical differences between type Ⅲ and other types (P<0.05). For the distance of AE, there were statistical differences between type Ⅲ and typeⅠ(P<0.05). Meanwhile, for the distance of AB, AC, AD, AE, AF, AG, there were statistical differences between male and female (P<0.05). For the distance of  β, AB, there were statistical differences between right and left (P<0.05).    Conclusion  Under the β, the sustentaculum tali is classified into three types, with the typeⅡas the main type. When treating calcaneal fractures with internal fixation, direction of screw implant can choose between 70° to 80° as soon as possible. The morphology and classification of sustentaculum tali are of certain clinical implication to treat calcaneal fractures. References | Related Articles | Metrics

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Comparison on the establishment of proprioceptive hypoesthesia model of anterior cruciate ligament by open surgery and minimally invasive surgery ZHANG Lei FENG Qi WANG Ping CHENG Han-wen XIONG Lu-jing 2022, 53 (4):  520-525.  doi: 10.16098/j.issn.0529-1356.2022.04.016 Abstract ( )   PDF (6156KB) ( )   Objective  To provide an optimized animal model for basic research by comparing the establishment of proprioceptive hypoesthesia model of anterior cruciate ligament (ACL) by open surgery and minimally invasive surgery.    Methods  Totally 30 normal cynomolgus monkeys were randomly divided into five groups: minimally invasive surgery group: unilateral ACL injury under arthroscope, n=6; open surgery group: unilateral ACL injury through direct incision of knee joint, n=6; minimally invasive sham operation group: unilateral ACL without injury through arthroscopic cleaning only, n=6; open surgery group: unilateral ACL exploration without injury through direct incision of knee joint, n=6; positive normal group: no surgical intervention, n=6. Four weeks later, the neurophysiological examination ［somatosensory evoked potentials (SEPs) and motor nerve conduction velocity (MCV) ］ was carried out. The macaque was killed and its unilateral ACL was stained with gold chloride. The number and variation of ACL proprioceptors were observed and recorded.    Results  In the aspect of neuroelectrophysiology, except the normal group, the latency of SEPs and MCV were prolonged, and the amplitude decreased. In terms of proprioceptors, the total number of minimally invasive surgery group: 578.00±12.68, amplitude: 36.33±3.72; total number of open surgery group: 367.67±9.33, amplitude: 77.00±5.55; total number of open sham operation group: 969.00±18.26, amplitude: 0±0; total number of normal group: 970.46±16.34, amplitude: 0±0, the total number of proprioceptors decreased and the number of variation increased in all four groups except the normal group. At the same time, in the comparison of the two aspects, there were significant differences between the open operation group and the minimally invasive operation group, the open operation group and the open sham operation group, and the minimally invasive operation group and the minimally invasive sham operation group (P<0.05). There were no significant difference between the open sham operation group and the minimally invasive sham operation group, and between the two groups and the normal group, respectively (P>0.05).     Conclusion  Both the open operation group and the minimally invasive operation group can build ACL proprioceptive hypoesthesia model, but the minimally invasive operation has less damage to the tissues around ACL, more scientific and single model, and less experimental error, which is of great significance in the basic research of ACL injury. References | Related Articles | Metrics

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Dicer requiring for astrocyte quiescence in developing brains LIU Kun CHEN Jing-fei WANG Shou-yu LI Tao GAO Xing YU Bin MEI Feng XIAO Lan 2022, 53 (4):  407-411.  doi: 10.16098/j.issn.0529-1356.2022.04.001 Abstract ( )   PDF (12011KB) ( )   Objective  To explore whether microRNAs(miRNA) in astrocytes participate in regulating the phenotypic switch in the developing central nervous system(CNS).     Methods  The hGFAP-CreERT;Dicer fl/fl;mT/mG and Dicer fl/fl;mT/mG mice were generated and induced by tamoxifen(TMF). They were divided into 7 days, 10 days, 14 days Dicer conditional knock out (Dicer CKO) group and control group according to their age and the expression of hGFAP-CreERT. These 24 mice were divided into six groups and each group contained four mice. Then the change of astrocytes in the developing CNS was observed by immunofluorescenct staining.     Results  Both of the densities of astrocytes and microglial cells  increased in the Dicer CKO mice brains as compared to littermate controls.      Conclusion  Dicer and miRNA of astrocytes are important for astrocytes quiescence in developing brains. References | Related Articles | Metrics

Cell and Molecules Biology

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Association between the polymorphism and expression level of the family microRNA-181 and susceptibility of ischemic stroke LEI Ming TAN Tan HUANG Yan-yun WEI Ye-sheng 2022, 53 (4):  470-478.  doi: 10.16098/j.issn.0529-1356-2022.04.010 Abstract ( )   PDF (3124KB) ( )   Objective  To explore the association between rs16927589, rs77418916, rs8108402 of the family microRNA(miR)-181 polymorphisms and ischemic stroke(IS)，and compare the expression of miR-181 genes between control group and IS group, further explore the association between polymorphisms and the expression levels of genes, to provide assistance for the prevention and treatment of IS.    Methods  SNaPshot technique and DNA sequencing were used to examine the single nucleotide polymorphism(SNP) genotypes of 349 patients of IS and 372 controls, serum lipid level was detected by biochemical analyzer 7600; The expression level of miR-181 genes in peripheral blood mononuclear cells of control group and IS group were detected by ABI7500 Real-time PCR.    Results  The genotype and allele of rs8108402 were compared between the control group and IS group, and it was found that compared with CC genotype, the risk of IS was significantly increased among people with CT genotype, TT genotype was opposite ［CC vs CT：odds ratio(OR)=1.56，95% confidence interval(CI),1.11-2.18,P=0.011;CC vs TT: OR=0.25, 95%  CI,0.10-0.62, P=0.003］. However, there were no correlation with IS of rs16927589 and rs77418916 polymorphisms. Stratified analysis of rs8108402 showed that the low-density lipoprotein (LDL-C) of IS patients with CC genotype was higher than that of IS patients with CT genotype (P<0.05), and the polymorphism of rs8108402 might be correlated with the clinical manifestations of IS. In IS group, the expression of miR-181a, miR-181b, miR-181c were obviously higher than that in control group, the difference was statistically significant (P<0.05), while the expression of miR-181d significantly lower than the control group, but the difference was not statistically  significant (P>0.05). Further analysis of rs8108402 polymorphism and gene expression level showed no correlation between rs8108402 polymorphism and gene expression level.     Conclusion  The CT and TT genotypes at rs8108402 of miR-181c increase the risk of IS, while the CTC haplotype increase the risk of IS. The polymorphism of rs8108402 is correlated with the level of LDL-C. There are significant differences in the expression of miR-181 gene clusters between the normal control group and the IS group, and miR-181 clusters may be potential predictive targets and therapeutic targets of IS. References | Related Articles | Metrics

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Effect and mechanism of bromodomain-containing protein 4 inhibitor JQ1 on formaldehyde solution induced inflammatory pain in mice CHEN Ling LIU Zhi-wen YANG Bin ZHOU Xin XIE Zhi-yan ZENG Jia-yu ZHAO Ke-xin CAO Wen-yu ZHONG Xiao-lin 2022, 53 (4):  412-417.  doi: 10.16098/j.issn.0529-1356.2022.04.002 Abstract ( )   PDF (4008KB) ( )   Objective  To explore the expression and role of bromodomain-containing protein 4(BRD4) in spinal cord of mice which suffered inflammatory pain induced by formaldehyde solution.    Methods  Thirty-two ICR mice were randomly divided into normal saline group and formaldehyde injection 5 minutes, 30 minutes and 60 minutes groups, with 8 mice in each group. The expression of BRD4 protein and mRNA in spinal cord of mice in each group were detected by Western blotting (n=4/group) and Real-time PCR (n=4/group); 66 mice were randomly divided into formaldehyde injection group, vehicle (DMSO) plus formaldehyde injection group and 6.25, 12.5, 25 and 50 mg/kg JQ1 injection plus formaldehyde solution group, with 11 mice in each group. The effect of BRD4 inhibitor JQ1 on spontaneous pain in each group was observed (n=11/group); Immunohistochemistry (n=3/group), Real-time PCR (n=4/group) or Western blotting (n= 4/group) were used to detect the effects of 25 mg/kg JQ1 on the expression of c-fos and glutamate receptor 2(GluR2) in the spinal cord of model mice.    Results  The result  showed that levels of BRD4 protein (P<0.01) and mRNA in spinal cord increased significantly 30 min and 60 min after formaldehyde solution injection (P<0.05). The behavioral test showed that both 25 mg/kg and 50 mg/kg JQ1 administration could reduce the second phase spontaneous pain compared with the solvent (DMSO) group (P<0.05). Furthermore, immunohistochemistry and Real-time PCR result  showed that 25 mg/kg JQ1 injection could significantly reduce positive numbers (P<0.01) and high mRNA expression of c-fos in mouse spinal cord induced by formaldehyde solution (P<0.05), and the Western blotting result  showed that 25 mg/kg JQ1 administration could significantly reduce the expression of glutamate receptor GluR2 (P<0.001).     Conclusion  BRD4 may play an important role in the induction of central sensitization of inflammatory pain, and JQ1 may alleviate inflammatory pain behavior by inhibiting the formation of central sensitization of pain. References | Related Articles | Metrics

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Effect of draxin knockdown on the projection of 23C10-positive neural fibers in the embryonic chick hindbrain WANG Zi-yi ZHANG Shu-han ZHANG Zi-ren ZHANG Wen-wen HU Ya-nan ZHANG San-bing SU Yu-hong 2022, 53 (4):  418-423.  doi: 10.16098/j.issn.0529-1356.2022.04.003 Abstract ( )   PDF (1515KB) ( )   Objective  To investigate the effects of the downregulation of draxin expression on the projection characteristics of 23C10-positive neural fibers in the chick embryonic hindbrain.    Methods  The vitro incubation of HH stages 21-22 chick embryonic hindbrain biopsy with alkaline phosphatase （ALP）protein was used as control group. The incubation of HH stages 21-22 chick embryonic hindbrain biopsy with draxin-ALP fusion protein was used as experimental group. The number of embryonic hindbrain for each group was 10. To detect whether 23C10-positive neural fibers could directly bind to draxin protein or not；In ovo electroporation using empty vector in the chick embryonic hindbrain was used as control group. In ovo electroporation with small interfering RNA(siRNA) expressing vector for reducing draxin expression in the chick embryonic hindbrain was used as experimental group. The number of embryonic hindbrain for each group was 18. The effect of the down-regulation of draxin expression and the change of projection characteristics of 23C10-positive neural fibers were observed to check whether the down-regulation of draxin expression would affect the distribution of 23C10-positive fibers.   Results   Most portion of draxin protein could overlap with 23C10-positive neural fibers in HH stages 21-22 chick  embryonic hindbrain biopsies；After expression of the siRNA plasmid against draxin by electroporation, the expression level of draxin protein was significantly reduced, and the distribution of 23C10-positive fibers was scattered in the dorsal hindbrain on the electroporated side at HH stages 25-26 of chick embryos（P<0.05）.     Conclusion  Draxin protein may directly bind to 23C10-positive fibers in hindbrain, and it plays an important regulatory role in the fasciculation of 23C10-positive fibers during chick embryonic development. References | Related Articles | Metrics

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MicroRNA-486 targeting TRIM10 to inhibit Parkinson’s disease cell model damage LIAO Jian-yun JIANG Xin-liu XIE Wei-fa 2022, 53 (4):  424-431.  doi: 10.16098/j.issn.0529-1356.2022.04.004 Abstract ( )   PDF (7709KB) ( )   Objective  To study the effect and mechanism of microRNA-486(miR-486) on 1-methyl-4-phenylpyridine (MPP+)-induced apoptosis of Parkinson’s disease (PD) PC12 cells in vitro.    Methods The experiment was divided into control group, PD (MPP+ induced PC12 cells), miR-NC (transfected mimics control, MPP+ induced PC12 cells), miR-486 group (transfected miR-486 mimics, MPP+ induced PC12 cells), miR-486+vector group (co-transfected miR-486 mimics, pcDNA3.1), miR-486+TRIM10 group (co-transfected miR-486 mimics, pcDNA3.1-TRIM10)，n=9 each group. CCK-8 method  was used to analyze cell proliferation changes, flow cytometry was used to analyze cell apoptosis levels, Western blotting was used to analyze changes in Bax and Bcl-2 protein expression, thiobarbituric acid method  was used to detect malondialdehyde (MDA) content in cells, fluorescence probe method  was used to detect the level of reactive oxygen species (ROS) in the cells, and 2, 4-dinitrophenylhydrazine was used to analyze the level of lactate dehydrogenase (LDH) in the culture supernatant. Bioinformatics software was used to predict the target genes of miR-486, and the detection of targeting relationship between imiR-486 and TRIM 10 by dual luciferase reporter gene assay.   Results  Compared with the control group, the cell survival rate and Bcl-2 protein expression in the PD group decreased, while the apoptosis rate, Bax protein expression, MDA, ROS and LDH levels increased. Compared with the miR-NC group, the cell survival rate and Bcl-2 protein expression in the miR-486 group were increased, and the apoptosis rate, Bax protein expression, MDA, ROS and LDH levels were decreased. MiR-486 targeted down-regulation of TRIM10 expression. Compared with the miR-486+vector group, the miR-486+TRIM10 group decreased the cell survival rate and Bcl-2 protein level, while the apoptosis rate, Bax protein level, MDA, ROS and LDH levels increased.  Conclusion  Up-regulation of miR-486 targeted and inhibited TRIM10 to reduce MPP+induced apoptosis in in vitro Parkinson’s PC12 cell models. References | Related Articles | Metrics

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Anti-oxidative stress mechanism of velvet antler polypeptide in Alzheimer’s disease model mice LANG Wei-ya ZHANG Chun-mei ZHANG Yu-di LIU Zhong-jin YI Tong-hui ZHANG Ke-shuang ZHANG Hai-yan 2022, 53 (4):  432-439.  doi: 10.16098/j.issn.0529-1356.2022.04.005 Abstract ( )   PDF (6239KB) ( )   Objective  To study the effect of velvet antler polypeptides (VAP) on antioxidant in Alzheimer’s disease model mice.    Methods  Eight months old male amyloid precursor protein (APP)/presenilin-1 (PS1) double transgenic mice were selected as Alzheimer’s disease (AD) model and divided into the model group and the VAP intervention group，12 in each group．Besides，normal mice of the same brood (with no transgene) were recruited as a control group (n=12) ．After 6 months of intragastric administration, behavior, morphology and oxidative stress related indicators were detected.SH-SY5 cells were used to establish AD model of damaged by Aβ25-35. The expression levels of APP and β-secreatase-1(BACE1) protein in mouse hippocampus were detected by Western blotting. VAP intervention group SH-SY5Y cells was cultured with VAP (500g/L) and amyloid  β(Aβ)25-35(25 μmol/L) for 24 hours. Control group cells were normally cultured by DMEM medium. Cell apoptosis, membrane potential, reactive oxygen species (ROS) levels and oxidative stress related indexes were detected.    Results  In animal models, compared with the model group, the escape latency of mice in the VAP intervention group was shortened (P＜0.05). The neuronal cells in the CA1 region of the hippocampus of the model group were reduced and arranged disorderly. The arrangement of the VAP intervention group was relatively regular, and the morphology was significantly improved. Compared with the model group, senile plaques were decreased in the VAP intervention group. Compared with the model group, the malondialdehyde (MDA) content of the VAP intervention group increased, and the superoxide dismutase（SOD）and glutathione peroxidase (GSH-Px) content increased, the difference was statistically significant. Compared with the control group, the APP and BACE1 content in the model group increased. Compared with the model group, the contents of APP and BACE1 in the VAP intervention group decreased, and the difference was statistically significant (P＜0.05). In the cell model, the apoptosis rates of the VAP intervention group decreased. Compared with the model group, the mitochondrial membrane potential of the VAP intervention group increased, the content of ROS decreased, the content of MDA decreased, and the content of SOD and GSH-Px increased. The difference were statistically significant (P＜0.05).     Conclusion  VAP has a protective effect on oxidative stress damage caused by Alzheimer’s disease model animals and cells, which may be achieved by reducing ROS production and increasing the activity of antioxidant enzymes to reduce Aβ deposition.  References | Related Articles | Metrics

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Relationship between nucleoprotein TAR DNA/RNA binding protein 43 and mouse amyotrophic lateral sclerosis TANG Wei-bo LIU Li SHEN Jing-ling 2022, 53 (4):  440-446.  doi: 10.16098/j.issn.0529-1356.2022.04.006 Abstract ( )   PDF (14587KB) ( )   Objective  Transgenic mice expressing human TAR DNA/RNA binding protein 43 (hTDP-43) mutant protein in spinal cord motor neurons were constructed using HB9 promoter to establish a disease model of amyotrophic lateral sclerosis (ALS) and explore the mechanism of ALS induced by hTDP-43 mutation.    Methods  HB9 promoter junction mutant hTDP-43 vector was constructed in vitro, and the positive transgenic mouse strains were  prepared by prokaryotic injection and screened(There were 8~10 mutations at Q331K and M337V). Gait analysis, rotary rod fatigue test, and suspension test were used to detect locomotion ability of mice. Immunohistochemistry, immunofluorescence staining and Western blotting were used to detect hTDP-43, phosphorylated HTDP-43 (p-hTDP-43), Caspase-3, cleaved Caspase-3, respectively. Expression of ubiquitin, β-tubulinⅢ(Tuj1), Ki67 and cyclin-dependent kinase 5 (CDK5)proteins were also detected.    Results  In transgenic mice expressing mutant hTDP-43 protein in spinal motor neurons, both hind limbs were atrophied to the trunk side, and motor function showed progressive decline with increasing age. hTDP-43, p-hTDP-43, Caspase-3, and cleaved Caspase-3 were observed in spinal motor neurons Caspase-3 positive staining and ubiquitin protein positive inclusion body, and in vitro isolation and culture of spinal motor neurons, it was found that hTDP-43- and ubiquitin protein co-located in choline acetyl translocation enzyme (ChAT）positive motor neurons, accompanied by ectopic expression of CDK5.     Conclusion  The mutant HDP 43 protein expressed in mouse spinal cord motor neurons can promote the re-entry of differentiated mature neurons into the cell cycle, leading to the occurrence of ALS. References | Related Articles | Metrics

Cancer Biology

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Reversing malignant biological behavior of osteosarcoma by knocking down 3-phosphoglycerin dehydrogenase targeting energy metabolism  ZHOU Hong KANG Quan XIE Sheng-nan CHEN Jie SHI Yu-lu LUO Qing 2022, 53 (4):  488-497.  doi: 10.16098/j.issn.0529-1356.2022.04.012 Abstract ( )   PDF (25131KB) ( )   Objective  To investigate the effect of knock-down 3-phosphoglycerin dehydrogenase (PHGDH) targeting energy metabolism on malignant biological behavior and osteogenic differentiation of human osteosarcoma 143B cells.    Methods  Real-time PCR and Western blotting were used to detect the expression of PHGDH in osteoblasts hFOB1.19 and osteosarcoma cells TE85, MG63 and 143B with different malignant degrees. The short hairpin RNA(shRNA)-PHGDH recombinant plasmid was transfected into 143 B cells by liposome transfection method. The expression of PHGDH was detected by Real-time PCR and Western blotting. Crystal violet staining, cell counting and CCK-8 assay were used to detect cell proliferation; wound healing assay was used to detect cell parallel migration ability, and Transwell assay was used to detect cell vertical migration and invasion ability. Annexin V-FITC/PI double staining and DAPI staining were used to detect apoptosis; Alkalinephosphatase(ALP) staining and alizarin red S staining were used to detect osteogenic differentiation. Western blotting was used to detect the expression of Runt related transcription factor 2 (Runx2) and osteocalcin (OC). The expression of genes related to energy metabolism, glucose  transporter-1(GLUT1), 6-phosphofructokinase-1(PFK1), pyruvate kinae subtype M2(PKM2), lactate dehydrogenase A(LDHA) was detected by Real-time PCR. Lactic acid secretion was detected by lactic acid detection kit. Adenosine triphosphate(ATP) production was detected by ATP detection kit.   Results  The expression of PHGDH in 143B cells was significantly higher than that in hFOB1.19, MG63 and TE85 cells (P<0.01). After the transfection of shRNA-PHGDH recombinant plasmid, the expression of PHGDH in 143 B cells decreased (P<0.01), proliferation ability decreased (P<0.01), cell migration and invasion ability decreased (P<0.01), apoptosis rate increased (P<0.01), ALP staining positive rate increased (P<0.01), alizarin red staining positive rate increased (P<0.05), Runx2 (P<0.05) and OC expression increased (P<0.01), expression of genes related to energy metabolism (GLUT1,PFK1,PKM2,LDHA) decreased (P<0.01), lactic acid decreased (P<0.01), ATP increased (P<0.05).   Conclusion  Knocking down of PHGDH can inhibit the proliferation, migration and invasion of human osteosarcoma 143B cells through energy metabolism, promote their apoptosis and promote their osteogenic differentiation. References | Related Articles | Metrics

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PI3K inhibitor ZSTK474 enhancing the antitumor of vesicular stomatitis virus Δ51 against osteosarcoma LIU Li-zhu LI Chao-yi LIN Shi-wei QU Ye TANG Qiang 2022, 53 (4):  498-506.  doi: 10.16098/j.issn.0529-1356.2022.04.013 Abstract ( )   PDF (16370KB) ( )   Objective  To explore whether PI3K inhibitor combined with oncolytic virus can play an effective oncolytic effect on osteosarcoma.      Methods  The cytotoxicity to tumor cells was detected by MTT method, and the mechanism of enhancing the anti-tumor activity was explored by observation of the swelling of endoplasmic reticulum using electron microscope and the expression of apoptosis-related proteins using Western blotting. The tumor clearance ability of the combination of the PI3k inhibitor ZSTK474 and vesicular stomatitis virus Δ51 (VSVΔ51) was verified by anti-tumor experiment  in vivo. The apoptosis of tumor cells was verified by immunohistochemistry.      Results  PI3K inhibitor could be used as sensitizers of oncolytic VSVΔ51, and confirmed that they promoted the strong apoptosis of osteosarcoma cells by aggravating the stress of endoplasmic reticulum in tumor cells (P＜0.01). In vivo experiments also showed that PI3K inhibitors combined with VSVΔ51 could significantly promote the oncolytic effect of osteosarcoma (P＜0.001), and this combination therapy enhanced the infiltration of immune cells in the tumor (P＜0.001).       Conclusion  PI3K inhibitors combined with oncolytic virus is a potential therapy for osteosarcoma. References | Related Articles | Metrics

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Differential expression and bioinformatics analysis of epsin3 in colorectal cancer XIANG Li-chun JIANG Zhong-xiang JIANG Xiao-ye CHEN Xue-nuo JIANG Zheng 2022, 53 (4):  507-514.  doi: 10.16098/j.issn.0529-1356.2022.04.014 Abstract ( )   PDF (3709KB) ( )   Objective To investigate the expression and significance of the adaptor protein EPN3 (Epsin 3) in colorectal cancer in order to provide reference for further study of EPN3. Methods We used tools of GEPIA and GEDS to analyze the expression of EPN3 in colorectal cancer tissues and cells. SMART and cBioPortal databases were used to analyze the relationship between EPN3 gene methylation and copy number variation and its expression level. Metascape was used to complete analysis of gene ontology (GO) functional annotation and related pathways of EPN3 related genes and BioPlex was applied to constructed a protein network in HCT116 cell. We collected 13 pairs of colorectal cancer adjacent tissue and cancer tissue specimens, and detected EPN3 mRNA expression by using RT-qPCR. We observed the effect of abilities of cell proliferation, clone formation and migration via silencing EPN3 in HCT116 and HT29. Results GEPIA, GEDS, SMART and cBioPortal analyses showed that EPN3 was highly expressed in colorectal tumor tissues (p<0.01), and was related to methylation and copy number variation. The enrichment results of EPN3 related genes showed that it was mainly related to cell adhesion. And a protein interaction network constructed by CCDC130, TNFAIP1, PHGDH, EPN2, etc. was related to protein ubiquitination. RT-qPCR result showed that EPN3 was highly expressed in tumor tissues (p<0.05). Silencing EPN3 inhibited the proliferation, clone formation and migration abilities of HCT116 and HT29 cells. Conclusion EPN3 is highly expressed in colorectal cancer tissues and is related to cell adhesion and protein ubiquitination. Down-regulated EPN3 can inhibit abilities of proliferation, clone formation and migration of HCT116 and HT29 cells, and this could provide a reference for further research on EPN3. References | Related Articles | Metrics

Anthropology

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Comparison of head and face measurement traits in Chinese Han, Li, Uygur and Tibetan population LU Jiu-zheng QIAO Hui SUN Chang TAN Jing-ze 2022, 53 (4):  526-533.  doi: 10.16098/j.issn.0529-1356.2022.04.017 Abstract ( )   PDF (1465KB) ( )   Objective  To investigate the relationship between the head and face traits of Han, Li, Uygur and Tibetan population.    Methods  From 2008 to 2011, sixteen head and face measurement traits were measured in Chinese young individuals from Jiangsu, Hainan, Xinjiang and Tibet, with sample size of 1034 ( 348 Han, 165 Li, 305 Uygur, 216 Tibetan ). The gender difference analysis, commonalities and differences between populations and principal component analysis were carried out.    Results  Most of the sixteen head and face measurement traits showed significant gender differences, the mean measurements of most features of males were bigger than those of females in four populations. Samples of the four populations usually had brachycephaly and hyperbrachycephaly. Most Han, Li and Uygur belonged to euryprosopy. Han, Li and Tibetan were typically mesorrhiny. The homogenous features of head and face of the four populations were shown. However, the head and face of the four populations also had some heterogeneous features. For instance, Han had the broadest face and Li had the broadest nose among all four populations. Furthermore, Uygur had the narrowest nose and Tibetan had the narrowest face. The result  in population relationships presented that the head and face measurement traits of Han and Li were relatively similar, while those of Uygur and Tibetan were narrower.     Conclusion There are commonalities and differences in the head and face measurement traits among the four populations. The group relationship between Han and Li is relatively similar, and that between Uygur and Tibetan is relatively close. The homogeneity and heterogeneity between populations can provide data and theoretical support for genetics, forensic science and other fields.  References | Related Articles | Metrics

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Heath-Carter somatotype of Tajik adults YU Ting NUERZHATI Sha-di-ke WAISIMAI Abuduke-li-mu ZILALA Maimaiti-tuer-sun TUOHETIGULI Maimai-ti-ming MAHERE Tuer-hong LI Xin WEN You-feng 2022, 53 (4):  534-539.  doi: 10.16098/j.issn.0529-1356.2022.04.018 Abstract ( )   PDF (971KB) ( )   Objective  To explore the somatotype characteristics and changing rules of Tajik adults.    Methods  The Heath-Carter body type method  was used to determine the body size of 280 (124 males and 156 females) Tajik adults.    Results  The average body size of Tajik males and females were 4.3-3.1-1.8 and 7.0-3.1-1.1, respectively, and both are mesomorphic endomorphy.The ectomorphy of Tajik nationality were negatively correlated with age, female endomorphy and mesomorphy were positively correlated with age, while endomorphy and mesomorphy were not correlated with age. With increasing age, the difference in body shape between female age groups was more obvious than that of males.     Conclusion The Tajik have less skeletal muscle mass, and women have developed body fat, which is different from the Tibetan people and other people in the Altaic language family. References | Related Articles | Metrics

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Hypertension status and its biochemical indexes analysis in Zhuang-Dong ethnic groups YI Can YU Hui-xin LI Yong-lan 2022, 53 (4):  540-546.  doi: 10.16098/j.issn.0529-1356.2022.04.019 Abstract ( )   PDF (810KB) ( )   Objective  To investigate the prevalence rate and biochemical characteristics of hypertension in Zhuang-Dong ethnic groups in China.    Methods  Blood pressure and biochemical indexes were measured in 7411 cases (3248 males and 4163 females) from 5 ethnic groups of Zhuang-Dong nationality.    Results  The difference of crude prevalence rate of hypertension among Zhuang-Dong language groups was statistically significant. The crude prevalence of hypertension in males of the same age group was higher than that in females. Systolic blood pressure and diastolic blood pressure in males and females increased with age, and the crude prevalence of hypertension also increased with age. The mean values of triglyceride, cholesterol and low density lipoprotein in males of Zhuang-Dong language group were all lower than those of females. The mean levels of blood uric acid, blood urea and blood lipid in hypertensive patients were significantly higher than those with normal blood pressure; the mean levels of bilirubin and serum protein in hypertensive patients were close to those with normal blood pressure; the mean levels of blood lipid in hypertensive patients were more than the normal range.     Conclusion  The prevalence of hypertension in Zhuang-Dong ethnic group is high. The patients with hypertension should pay attention to the increase of blood uric acid, blood urea and blood lipid. References | Related Articles | Metrics