Objective  To investigate the effects of neuroligin-1, -2 (NLGN-1,-2) on oligodendrocyte(OLs) differentiation and myelination in the central nervous system.   Methods  OLs were cultured in vitro in the presence of different concentrations of NLGN-1 and NLGN-2. Morphological differentiation of OLs was observed by immunofluorescent staining and mRNA expression levels of myelin-associated genes were detected by Real-time PCR. Western blotting was used to detect the expression of myelin-related proteins. Results  NLGN-1, -2 accelerated the differentiation of oligodendrocyte precursor cells (OPCs) into mature OLs, and promoted the ability of myelin sheath formation. In vitro culture conditions, the dosage of 500 μg/L had the best promotion effect on OLs differentiation and maturation, and NLGN-2 had better promoting effect than that of NLGN-1. Furthermore, the mRNA expression levels of myelin-associated genes myelin protein P0(MPZ), myelin basic protein(MBP) increased after the neuroligins treatments detected by  Real-time PCR. Western blotting result  showed that the expressions of MBP and MPZ increased significantly after 500 μg/L treatment with NLGN-1 and NLGN-2 for 12 hours. Conclusion  NLGN-1, -2 promote OLs differentiation and myelination. The positive effect of NLGN-2 is greater than that of NLGN-1 significantly, suggesting that the treatment with inhibitory synaptic-associated cytokines may improve the ability of myelin sheath formation in the central nervous system.

Objective To explore the effect of scutellarin on lipopolysaccharide (LPS) induced neuroinflammation in BV-2 microglia cells.   Methods  BV-2 microglia were cultured and randomly divided into 6 groups: control group (Ctrl), cyclic GMP-AMP synthetase（cGAS）inhibitor RU320521 group (RU.521 group), LPS group, LPS+RU.521 group, LPS+ scutellarin pretreatment group (LPS+S) and LPS+S+RU.521 group. The expressions of cGAS, stimulator of interferon gene (STING), nuclear factor kappa B (NF-κB), phosphorylated NF-κB (p-NF-κB), neuroinflammatory factors PYD domainscontaining protein 3（NLRP3) and tumor necrosis factor α（TNF-α）in BV-2 microglia were detected by Western blotting and immunofluorescent double staining (n=3).   Results Western blotting and immunofluorescent double staining showed that compared with the control group, the expression of cGAS, STING, p-NF-κB, NLRP3 and TNF-α in BV-2 microglia increased significantly after LPS induction(P＜0.05), while the expression of cGAS, STING, p-NF-κB, NLRP3 and TNF-α in LPS+S group were significantly lower than those in LPS group (P＜0.05). Treatment with cGAS pathway inhibitor RU.521 showed similar effects as the pre-treatment group with scutellarin. In addition, the change of NF-κB in each group was not statistically significant（P＞0.05）.   Conclusion Scutellarin inhibits the neuroinflammation mediated by BV-2 microglia cells, which may be related to cGAS-STING signaling pathway.

Objective To investigate the protective effect and mechanism of acellular nerve allografts (ANA) combined with electroacupuncture on spinal ganglia in rats with sciatic nerve injury (SNI).   Methods Totally 50 male adult SD rats were randomly selected for this experiment. Ten rats were prepared for the ANA. Forty male SD rats were randomly divided into normal group, model group, ANA group and combinational group, with 10 rats in each group. The SNI model was established by cutting off the nerves 10 mm at the 5 mm on the inferior border of piriformis after separating the right sciatic nerves. The rats in the ANA group were bridged with ANA to the two broken ends of injured nerves. The rats in the combinational group were treated with electroacupuncture 2 days after ANA bridging, Huantiao (GB30) and Yanglingquan (GB34) were performed as the acupuncture points, each electroacupuncture lasted 15 minutes and 7 days as a course of treatment, 4 courses in all. Sciatic nerve conduction velocity was measured by electrophysiology to evaluate the regeneration of damaged axons. Morphology of spinal ganglia was observed by Nissl staining. The expression of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) were detected by Western blotting and immunofluorescent staining.   Results Compared with the normal group, the sciatic nerve conduction velocity in model group decreased significantly (P<0.01), Nissl bodies in neurons of spinal ganglia were swollen and dissolved, with incomplete structure and the number decreased dramatically (P<0.01), while the level of NGF and BDNF also decreased significantly(P<0.01). Compared with the model group, the sciatic nerve conduction velocity in ANA and combinational groups strongly increased (P<0.01), the damage of Nissl bodies in neurons of spinal ganglia reduced and the number obviously increased (P<0.01), the level of NGF and BDNF increased considerably (P<0.01). Compared with the ANA group, the sciatic nerve conduction velocity in combinational group increased significantly (P<0.01), the morphology of Nissl bodies in neurons of spinal ganglia were more regular and the number increased (P<0.01), moreover, the level of NGF also increased significantly (P<0.01).   Conclusion ANA combined with electroacupuncture can enhance the sciatic nerve conduction velocity, improve the morphology of neurons in spinal ganglia and play a protective effect on spinal ganglia. The mechanism can be related to the higher expression of NGF and BDNF proteins, especially the expression of NGF protein. 

Objective To observe the effect of catgut implantation at acupoint（CIAA）on the learning and memory function, hippocampal microangiogenesis, and the mRNA and protein expression of angiopoietin-1 (Ang-1)/vascular endothelial growth factor (VEGF) and its receptor TEK tyrosine kinase(TIE2)/VEGF receptor 2(VEGFR2) in rats with vascular dementia (VD). To explore the mechanism of catgut implantation at acupoint in preventing and treating VD.   Methods Using a random number table, VD rats were divided into a model group, a nimodipine group, and an catgut implantation at acupoint group, and a sham operation group was set up, with 10 rats in each group. On the 7th day after surgery, the treatment groups were given catgut implantation at acupoint and nimodipine gastric lavage for 21 days. After treatment, Morris water maze behavioral test was performed. HE staining was used to observe hippocampal CA1 tissue. CD34 immunohistochemical staining was used to detect hippocampal microvascular density (MVD). Real-time PCR and Western blotting were used to detect the mRNA and protein expression of Ang-1/VEGF and its receptor TIE2/VEGFR2 in the hippocampus.   Results Compared with the model group, the average escape latency of the other groups was significantly shortened, and the target quadrant residence time was significantly prolonged (P<0.01, P<0.05). Compared with the model group, the number of nucleolus and well-formed pyramidal cells in hippocampal CA1 area of the catgut implantation at acupoint group and the nimodipine group increased in varying degrees, and they were arranged more closely, with only a few cells scattered and swollen. In the sham surgery group, a few CD34 positive cells were scattered. The treatment groups had more closely distributed CD34 positive cells with significant staining compared to the model group. The MVD of the model group was significantly higher than that of the sham surgery group (P<0.01). Both nimodipine group and catgut implantation at acupoint group had higher MVD than the model group (P<0.05, P<0.01). Compared with the sham surgery group, the mRNA and protein expression of Ang-1/VEGF and its receptor TIE2/VEGFR2 in the model group increased significantly (P<0.01, P<0.05).   Compared with the model group, both nimodipine group and catgut implantation at acupoint group had higher mRNA and protein expression of Ang-1/VEGF and its receptor TIE2/VEGFR2 (P<0.01, P<0.05). Conclusion  Catgut implantation at acupoint can improve the learning and memory abilities in VD rats, promote hippocampal microvascular angiogenesis, which may be related to the up-regulation of Ang-1/VEGF and its receptor TIE2/VEGFR2 mRNA and protein expression.

Objective To investigate the effect of microglia activation regulated by C-X3-C motif chemokine ligand 1 (CX3CL1)- C-X3-C motif chemokine receptor 1 (CX3CR1) pathway on memory function in hemorrhagic shock/resuscitation rats.   Methods The experiment was divided into two parts. In the first part, the rats were randomly divided into sham group, model-0.5 hour group, model-1.5 hour group, model-3 hour group, 10 rats in each group. There were differences in the time of hemorrhagic shock among each group. In the second part, rats were randomly divided into control group and CX3CL1 group, 10 rats in each group. The rats in CX3CL1 group were treated with CX3CL1 protein factor (intraventricular injection), and the rats in control group were treated with saline. All rats were trained in Morris water maze experiments before model construction, and tests of Morris water maze experiments were carried out after 4 days of model construction. After completion, the whole brains were taken for HE staining and immunohistochemical staining. Cerebrospinal fluid was taken for detection of inflammatory cytokines, and hippocampus tissues were taken for Real-time PCR detection and Western blotting detection.  Results Compared with the sham group, the escape latency of rats in model group increased, the number of platform crossings and the resident time in the third quadrant decreased. The neuronal state was impaired in HE staining in model group. In addition, compared with the sham group, the expression of ionized calcium binding adaptor molecule-1 (Iba1) in the brain of the rats in model group increased, the contents of tumor necrosis factor-α(TNF-α) and interleukin(IL)-6 in the cerebrospinal fluid increased, and the M1-type microglia markers CD16, TNF-α, IL-1β and inducible nitric oxide synthase (iNOS) mRNA content increased. At the same time, compared with the sham group, the expressions of CX3CL1 and CX3CR1 in the brain of model group decreased, and the expressions of phosphorylated nuclear factor-κB（p-NF-κB）and nucleotide binding oligomerization domain(NOD) -like receptor protein 3（NLRP3）increased. However, compared with the control group, rats in CX3CL1 group had reduced escape latency, increased platform crossing times and quadrant Ⅲ resident time, and recovered neuronal states. In addition, the expression of Iba1 in the brain of CX3CL1 group decreased, the contents of TNF-α and IL-6 in the cerebrospinal fluid decreased, the mRNA contents of M1-type microglia markers like CD16, TNF-α, IL-1β and iNOS decreased, and the mRNA contents of markers of M2-type microglia glial like CD206, transforming growth factor-β(TGF-β), arginase-1 (Arg1), Chitinase 3-like protein 1(Ym 1) increased.   Conclusion  CX3CL1 can help inhibit the excessive activation of microglia, induce the polarization of microglia to M2 type, inhibit the polarization of M1 type, reduce the release of inflammatory cytokines, and alleviate the memory function damage induced by hemorrhagic shock/resuscitation.

Objective  To discuss the relationship between activated glia cells in distal segment of the spinal cord and widespread pain.   Methods Fifty female rats were randomly divided into sham group, the chronic constriction injury of the infraorbital nerve (CCI-ION) group, CCI-ION + minocycline (Mino) group, CCI-ION + L-2-aminoadipic acid (LAA) group, and CCI-ION + normal saline (NS) group, n=10 for each group. CCI-ION model was established and Mino, LAA, and normal saline were delivered intrathecally to CCI-ION rats. Immunofluorescence staining was used to detect activated astrocytes and microglia in the medulla oblongata, cervical, thoracic, and lumbar spinal cord segments. On the 7th, 14th, 21st, 28th day, von Frey filaments were used to evaluate the mechanical withdrawal threshold of vibrissa pad, and electronic von Frey tactile pain meter was used to measure the mechanical withdrawal threshold of front paw, chest and hind paw. The radiant thermal stimulator was used to measure the thermal withdrawal threshold of hind paw.   Results After intrathecal injection of Mino to inhibit microglia, the activated microglia in each spinal cord segment decreased. Moreover, inhibiting astrocytes by using LAA significantly reduced activated astrocytes in spinal dorsal horn from distal segments. Behavioral assay showed that after intrathecal injection of Mino and LAA, the mechanical allodynia of vibrissa pad in CCI-ION rats was relieved. However, there was no significant difference (P>0.05)in the thermal and mechanical withdrawal thresholds in the hind paw of CCI-ION rats after intrathecal injection of Mino, while intrathecal injection of LAA significantly increased the thermal and mechanical withdrawal thresholds in the hind paw, indicating the relief of widespread pain induced by CCI-ION.   Conclusion The activated astrocytes in distal segments of the spinal cord mediated CCI-ION-induced widespread pain.

Objective To investigate the efeects of microRNA(miR)-103a-3p regulates tumor protein 53-regulated inhibitor of apoptosis 1(TRIAP1) on osteoblast differentiation and bone mass in ovariectomized mice.    Methods MC3T3-E1 cells were divided into normal group, miR-103a-3p-NC group, miR-103a-3p mimic group, miR-103a-3p mimic + TRIAP1-NC group, miR-103a-3p mimic + TRIAP1 mimic group. mRNA expression of miR-103a-3p, TRIAP1, P53 were detected by Real-time PCR; Cell proliferation and apoptosis were detected by MTT test and flow cytometry; cytoskeleton and mineralization of cells were detected by F-actin immunofluorescence staining and alizarin staining; alkaline phosphatase(ALP) activity was detected by ELISA. 24 female mice were divided into sham group, osteoporosis(OP) group, miR-103a-3p antagonist-NC group, miR-103a-3p antagonist group(six in each group), extract bilateral ovaries to establish an OP model, sham group mice only isolated fat around ovarian tissue. mRNA expression of miR-103a-3p, TRIAP1, P53, ALP, osteocalcin(OCN), osteopontin(OPN) of bone tissue were detected; microCT detect bone mineral density (BMD), bone mineral content (BMC); haematoxylin eosin staining was used to observe pathological changes of bone tissue.   Results After miR-103a-3p mimic was transfected into cells, the miR-103a-3p and P53 expression increased, TRIAP1 expression decreased, cell proliferation decreased, apoptosis increased, F-actin expression decreased, the number of calcium nodules decreased, and ALP enzyme activity decreased (P<0.01); however, after TRIAP1 mimic was additionally transfected into cells, the above result  caused by miR-103a-3p mimics were significantly reversed (P<0.01). In OP group, the miR-103a-3p and P53 expression in bone tissue increased, the TRIAP1, ALP, OCN and OPN expression decreased, BMD and BMC were decreased, and bone tissue construct was damaged(P<0.05); in miR-103a-3p antagonist group, the miR-103a-3p and P53 expression in bone tissue decreased, TRIAP1, ALP, OCN, OPN expression increased, BMD and BMC increased, and bone tissue construct was improved (P<0.05).   Conclusion MiRNA-103a-3p mediate TRIAP1/P53 to inhibit proliferation and mineralization of osteoblast, while miR-103a-3p antagonistic treatment reduce bone loss in OP mice.

Objective To investigate the association of 13 single nucleotide polymorphism (SNP) sites in 6 phalange-bone development related genes ［fibroblast growth factor receptor 2 (FGFR2), indian hedgehog signaling molecule (IHH), Msh homeobox 1(MSX1), Runx family transcription factor 2(RUNX2), SRY-box transcription factor 9(SOX9), Wnt family member 5A (WNT5A)］ with human index-ring finger length ratio (2D∶4D).   Methods Digital cameras were used to take frontal photographs of the hands of 731 college students (358 males and 373 females) in Ningxia, and image analysis software was used to mark anatomical points and measure finger lengths of index (2th) and ring (4th); genotyping of 13 SNP sites (rs1047057, rs755793, rs41258305, rs3731881, rs3100776, rs12532, rs3821949, rs45585135, rs3749863, rs1042667, rs12601701, rs1829556, rs3732750) for 6 genes by multiplex PCR; One-Way ANOVA or independent sample t-test indirectly assessed the association between 2D∶4D and 13 SNP sites. Results  Both left and right hand 2D∶4D were significantly higher in females than males in Ningxia college students (all P<0.01); no statistically significant differences in genotype and allele frequencies of the 13 SNP sites among different sexes (all P>0.05); among different sexes, male left hand 2D∶4D was significantly associated with the genotype of SOX9 gene rs12601701 site (P<0.05) and right hand 2D∶4D was significantly associated with the genotype of WNT5A gene rs1829556 site (P<0.05); the female right hand 2D∶4D was significantly associated with the MSX1 gene rs12532 (P<0.01) and rs3821949 (P<0.05) sites genotypes.  Conclusion SOX9 (rs12601701), WNT5A (rs1829556) and MSX1 (rs12532 and rs3821949) gene polymorphisms may be associated with the formation of 2D∶4D in Ningxia population.

Objective To explore the influence and mechanism of lentiviral vector-mediated steroidogenic factor-1 (SF-1) silence on decidual process of human endometrial stromal cells (hESCs).   Methods The endometrial tissues of patients with endometriosis (EM) and normal pregnant women were collected, and the differential expression of SF-1 was detected by Real-time PCR and immunohistochemical staining. hESCs were isolated from the endometrial tissue of normal pregnancy and identified, hESCs were divided into control group, estradiol (E2) + progesterone (P4) group, short hairpin RAN(shRNA, sh)-normal control(NC)+E2+P4 group, sh-SF-1+E2+P4 group, after the corresponding processing, the morphological changes of hESCs were observed under an inverted microscope. Real-time PCR and Western blotting were used to detect the mRNA expression levels and protein levels of human insulin-like growth factor-binding protein-1 (IGFBP-1) and prolactin (PRL) in cells, respectively, flow cytometry was used to determine the cycle distribution of the cells, immunofluorescence staining was used to observe the expression of the intracellular auto-bid microtubule-associated protein light chain 3(LC3), Western blotting was used to determine the protein level of intracellular autophagy-related proteins LC3-Ⅱ, LC3-Ⅰ and Beclin-1.   Results The relative expression of SF-1 mRNA and the positive rate of SF-1 protein in endometrial tissue of EM patients were higher than those of normal pregnancy endometrial tissue (P<0.05). Compared with the sh-NC+E2+P4 group, the cells in the sh-SF-1+E2+P4 group were mostly long-spindle, and there was no obvious decidualization. The relative mRNA expression and protein expression of IGFBP1 and PRL were significantly down-regulated (P<0.05), the proportion of cells in G0/G1 phase was significantly decreased, the proportion of cells in S phase was significantly increased (P<0.05), the ratio of LC3-Ⅱ/LC3-Ⅰ was significantly increased, and the relative expression of Beclin-1 protein was also significantly up-regulated (P<0.05).   Conclusion The expression of SF-1 in the endometrial tissue of EM patients is increased, and SF-1 silencing mediated by lentiviral vector can inhibit the decidualization process of hESCs, which may be related to regulating the level of autophagy.

Objective To determine the acetylation level of nucleophosmin (NPM) in female breast cancer and to discuss its function through mutation of modified lysine sites. To construct positive and negative NPM mutants on its acetylated lysine sites and to express them in breast cancer cells.   Methods Acetylation level and acetylated lysine sites of NPM in three breast cancer tissues and para-carcinoma tissues were detected by acetylome technology; NPM mutants were constructed by site-directed mutagenesis PCR, specific PCR products were digested by DpnI and transformed into Escherichia coli(E.coli) to obtain specific plasmids for mutants; The accuracy of mutants were verified by double restriction enzyme digestion and sequencing; The mutants were expressed in BT-549 cells by transient transfection and verified by RT-PCR method. Protein expression and acetylation level of NPM were validated by Western blotting; Function of NPM acetylation was analyzed by proteomic detection and bioinformatic analysis.   Results The 27th and 32nd lysine of NPM were highly acetylated in breast cancer tissues, which were 2.76 and 2.22 times higher than those in adjacent normal tissues, respectively; The NPM mutants showed the same molecular weight as that of wild type NPM and contained expected mutation sites; Corresponding NPM mRNA levels of BT-549 cells transfected with NPM mutants were significantly increased. With the increase of wild type NPM expression level, NPM acetylation level increased, while decreased after 27th lysine underwent negative mutation. NPM acetylation can significantly change the expression levels of 101 proteins in BT-549 cells, which are enriched in regulation of cellular macromolecule biosynthesis, DNA-template transcription, RNA biosynthesis and RNA metabolism process.   Conclusion NPM is highly acetylated in breast cancer and can play a key role in cellular macromolecule biosynthesis, DNA-templated transcription, RNA biosynthesis and RNA metabolism process. 

Objective To investigate the effects of epithelial transformation sequence 2 (ECT2) and p33ING1 on the metastatic activity of esophageal squamous cell carcinoma (ESCC) cells.   Methods The expressions of ECT2 and p33ING1 in esophageal squamous cell carcinoma tissues and adjacent tissues were detected by immunohistochemistry and Western blotting. Human esophageal squamous carcinoma cell line KYSE140 cells were divided into 4 groups: blank group, negative control (pcDNA 3.1 NC) group, overexpression group (pcDNA 3.1 ECT2) and inhibited expression group (si ECT2). MTT assay and cell colony formation assay were used to study the proliferation and growth ability of cells, Transwell assay and scratch assay used to study the invasion and migration ability of cells, and flow cytometry used to detect apoptosis and cell cycle, Western blotting used to detect the effect of ECT2 on p33ING1 protein.   Results ECT2 expression increased and p33ING1 expression decreased in esophageal squamous cell carcinoma tissues. Overexpression of ECT2 significantly increased the growth, colony formation, migration and invasion abilities of KYSE140 cells, and decreased the apoptosis rate and p33ING1 expression of KYSE140 cells. In addition, inhibition of ECT2 expression could reverse the above changes.   Conclusion The high expression of ECT2 can promote the growth and metastasis of esophageal squamous cell carcinoma KYSE140 cells and inhibit their apoptosis. The mechanism may be related to the inhibition of p33ING1 expression by ECT2.

Objective To analyze the factors associated with pain after arthroscopic rotator cuff bridge suture.   Methods According to the inclusion and exclusion criteria, the data of 112 patients with unilateral rotator cuff injury who received arthroscopic bridge suture in our department were collected and were investigated in the form of telephone follow-up. In this study, SPSS 23.0 was used to input data and conduct statistical analysis. Logistic regression analysis was used to analyze the correlation between the above influencing factors and postoperative pain.   Results A total of 112 patients were included for statistical analysis, single factor analysis revealed, including course of disease, smoking history, preoperative University of California, Los Angeles (UCLA) score, Constant score, numeric rating scale (NRS), size of rotator cuff tear, whether it was full-thickness tear and degree of tendon retraction might be related to postoperative pain (P<0.05). The age, gender, body mass index (BMI), drinking history, diabetes and hypertension were not related to postoperative pain (P>0.05). Multiple linear regression analysis concluded that there were four factors related to postoperative pain, and the correlation degree was preoperative NRS, preoperative UCLA score, tear size and smoking history.   Conclusion The causes of postoperative pain after arthroscopic rotator cauff repair are complex and diverse. Analyzing the cause of postoperative pain can effectively reduce the pain of patients and promote the recovery of shoulder joint function.

Objective To investigate the effects and mechanisms of peimine (PME) on chronic obstructive pulmonary disease (COPD) in mice.   Methods The mice were randomly divided into 4 groups (20 mice in each group), control group, PME group, chronic obstructive pulmonary disease group and treatment group. Animal models of COPD were induced in mice by lipopolysaccharide combined with smoke. The effects of PME on COPD model mice was analyzed by HE staining, transmission electron microscopy and the ratio of wet/dry weight of mouse lung tissue. The effects of PME on COPD model mice were analyzed by HE staining, transmission electron microscopy and the ratio of wet/dry weight of mouse lung tissue. The effects of PME on inflammatory factor tumor necrosis factor (TNF)-α, interleukin(IL)-6 and IL-1β in lung tissue were analyzed by ELISA and Western blotting. The effects of PME on oxidative stress in lung tissue were analyzed by dihydroethidium (DHE) staining and Western blotting. The effects of PME on nuclear factor kappa-B (NF-κB) pathway and nuclear factor erythroid 2-related factor 2(Nrf2) pathway were analyzed by protein immunoblotting.      Results Compared with the COPD group, PME treatment could significantly improve the lung tissue injury and the number of inflammatory cells in mice, and the wet/dry weight ratio of lung tissue was significantly reduced. Compared with the control group, the levels of TNF-α, IL-6 and IL-1β in the alveolar lavage fluid of COPD mice significantly increased, and the level of TNF-α, IL-6 and IL-1β in the alveolar lavage fluid of mice after PME treatment was significantly reduced. In addition, compared with the control group, the protein expression of TNF-α, IL-6 and IL-1β in the lung tissue of COPD mice significantly increased, and the level of TNF-α, IL-6 and IL-1β in the lung tissue of COPD mice after PME treatment were significantly reduced. Immunohistochemistry and Western blotting showed that the level of superoxide dismutase 2(SOD2) protein in COPD group was significantly lower than that in control group, while PME treatment could improve the level of superoxide dismutase protein. The analysis of MDA content in lung tissue showed that compared with the COPD group, the production of MDA in lung tissue of COPD mice was significantly inhibited after PME treatment. Protein Western blotting showed that PME treatment could prevent the phosphorylation of inhibitor of NF-κB(IκBα) and the transfer of NF-κB p65 to the cell nucleus, and the expression of Nrf2 and its main downstream target heme oxygenase-1(HO-1) in the lung tissue of mice treated with PME significantly increased.  Conclusion PME is able to inhibit inflammation and oxidative stress and improve lung tissues damage in the COPD model in vivo and this protection effect might be both the Nrf2 pathway activation and NF-κB pathway inhibition.

Objective To investigate the effects of microRNA (miR)-30a-regulated MAPK pathway on the formation of intercalation, inflammatory factors and vasoconstriction in a rat model of aortic coarctation.   Methods Fifty SD rats were selected to establish the rat model of aortic coarctation, and were randomly divided into control group, model group, miR-NC group, miR-30a group and miR-30a inhibitor group, 10 rats in each group. Histopathological changes in the aortic tissue and changes in the elastic fibers and collagen fibers of the aortic mesothelium were observed; The expression of miR-30a, systolic blood pressure before and after the intervention and the expression of serum inflammatory factors in each group were measured by PCR, tail artery manometry and ELISA; Matrix metalloproteinase (MMP)-6, MMP-2 protein expression and MAPK pathway were measured by Western blotting in each group. The expression of MMP-6, MMP-2 and MAPK pathway related proteins were measured by Western blotting.   Results The miR-30a inhibitor group improved the degree of vessel wall tearing and disorganized internal arterial wall arrangement; The miR-30a group improved vascular remodeling; miR-30a expression was higher in the model group compared with the control group, and lower in the miR-30a group and miR-30a inhibitor group compared with the miR-NC group, P<0.05; Before the intervention, the difference in systolic blood pressure between the groups compared was not statistically significant, P> 0.05; Compared with the control group, systolic blood pressure was higher in the model group, higher expression in the miR-30a group and lower expression in the miR-30a inhibitor group compared with the miR-NC group, P< 0.05; compared with the control group, tumor necrosis factor(TNF)-α, interleukin (IL)-6, IL-1β expression was higher in the model group, higher expression in the miR-30a group compared with the miR-NC group, lower expression in the miR-30a inhibitor group, P< 0.05; higher expression of TNF-α, MMP-6, MMP-2, Ras, Raf, P38MAPK, ERK1/2 proteins in the model group compared with the control group, higher expression in the miR-30a group compared with the miR-NC group, lower expression in the miR-30a inhibitor group, P<0.05.   Conclusion MiR-30a is involved in the process of aortic coarctation formation, inflammatory response, and regulation of aortic coarctation vascular remodeling, possibly through regulation of the MAPK signaling pathway.

Objective To investigate the effect of glucagon-like peptide 1 (GLP-1) receptor agonists exendin-4 on the secretion of cyclophilin A (CyPA) to inhibit atherosclerosis (AS) and vascular calcification in mice role of the process.   Methods Twenty ApoE-/- mice were randomly divided into model group and exendin-4 group, 10 mice in each group, and were fed with high-fat diet to establish AS model, another 10 wild-type C57BL/6J mice were taken as the control group, and the exendin-4 group was intraperitoneally injected with the GLP-1R agonist exendin-4, 1/d, for 8 weeks. After 8 weeks, the ELISA method  was used to determine the level of triglyceride(TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and CyPA, serum calcium level was detected by methylthymol blue colorimetric method, oil red O staining to detect the development of atherosclerotic plaques in the aorta, HE staining was used to observe the pathological changes of the aorta, Von Kossa staining was used to observe the calcium deposition in the aorta, immunohistochemical staining, Real-time PCR and Western blotting were used to detect the expression levels of aortic RUNX2 and bone morphogenetic protein 2（BMP-2）, immunofluorescent staining was used to detect the positive expression of CyPA in aortic tissue.   Results Compared with the control group, the serum levels of TG, TC, LDL-C, Ca and CyPA in the model group increased (P<0.05), the atherosclerotic plaque areas of the aorta increased (P<0.05), the aortic wall was thickened significantly and a large number of inflammatory cells were infiltrated, a large amount of calcium deposits were deposited in the aortic parietal membrane, the positive expression area ratio of RUNX2 and BMP-2, the relative mRNA expression of RUNX2 and BMP-2, the relative protein expression of RUNX2 and BMP-2 in aortic tissue all increased (P<0.05), and the red fluorescence of CyPA expression in aortic tissue was enhanced significantly. Compared with the model group, the serum levels of TG, TC, LDL-C, Ca and CyPA in the exendin-4 group decreased (P<0.05), the atherosclerotic plaque areas of the aorta decreased (P<0.05), the thickening of the aortic wall and the infiltration of inflammatory cells were alleviated significantly, the calcium deposition in the aortic wall was reduced, the positive expression area ratio of RUNX2 and BMP-2, the relative mRNA expression of RUNX2 and BMP-2, the relative protein expression of RUNX2 and BMP-2 in aortic tissue all decreased (P<0.05), and at the same time, the red fluorescence of CyPA expression in aortic tissue was weakened significantly.   Conclusion GLP-1 receptor agonists exendin-4 can inhibit atherosclerosis and vascular calcification in mice, and the mechanism may be related to the reduction of CyPA secretion.

Objective To analyze the antigen recognition sites of commercial and homemade antibodies against aquaporin (AQP) 9 ，and to identify the application effect.     Methods Western blotting was used to compare the efficacy of three commercial antibodies and self-made antibody in identifying AQP9 genotypes. The antigen recognition sites of four antibodies and their specificities in practical applications were analyzed.   Results Western blotting showed that protein bands of three commercial antibodies were detected in both WT and Aqp9-/- mice. The keyhole limpet hemocyanin(KLH) conjugated synthetic peptides corresponding to the three commercial antibodies were derived from rat, human and human, respectively. And The sequences of these three synthetic peptides were different from those of mice. AQP3/7 and AQP9 have similar molecular weight and were expressed in the liver with high homology. An obvious band of self-made antibody was observed at the 27 kD position in WT mice, but no band was observed at the corresponding position in Aqp9-/- mice.   Conclusion Commercial antibodies 1 and 3 can be used to assist in the identification of genotypes in Aqp9-/- mice. Homemade antibodies can accurately identify genotypes at the protein level.

Neurodevelopment and neuronal function are modulated by multiple factors including environment, genetics and epigenetics. As a post-translational modification, N-glycosylation is catalyzed by glycosyltransferase and involves in diverse biological processes. N-glycosylation is abundant in neuronal system, regulates the development and maturation of synapse, and inflammatory response of glial cells. The dysregulation of N-glycosylation induces neurological disorders including Alzheimer’s disease, congenital disorders of glycosylation, schizophrenia and epilepsy. In the present review, we have summarized the progresses of N-glycosylation in regulating neuronal and astrocytic function, and its roles in neurological disorders and related mechanisms. 

Myocardial infarction is one of the severe cardiovascular diseases. The patients with myocardial infarction die of heart failure or arrhythmia. In recent years, the studies in myocardial infarction therapies have advanced greatly, especially the preclinical experimental studies. The experimental studies of myocardial infarction often rely on animal models. Therefore, successful establishment of the myocardial infarction models has important application value in exploring the new techniques and measures for repairing the infarcted myocardium. In this paper, the techniques in establishment of the myocardial infarction models and strategies of their application are summarized.