Table of Content

2020, Volume 51 Issue 4
06 August 2020 Previous Issue    Next Issue
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Cancer Biology
Anatomy
Technology and Methodology
Review
Histology,Embryology and Developmental Biology
Neurobiology
Cell and Molecules Biology
Anthropology


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Cancer Biology

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Role of trefoil factor3/phosphatidylinositl 3-kinase/protein kinase B/nuclear factorκB in the proliferation and apoptosis of human papillary thyroid carcinoma cell line TPC-1 GUO Meng-yao SUN Lin LIN Xu ZHANG Wen-jing ZHANG Jing WU Jing-fang XUE Gang 2020, 51 (4):  528-535.  doi: 10.16098/j.issn.0529-1356.2020.04.009 Abstract ( )   PDF (2832KB) ( )   Objective To investigate the effects of trefoil factor 3 (TFF3) on the proliferation and apoptosis of human thyroid papillary carcinoma cell line (TPC-1) and its molecular mechanism.  Methods The lentiviral expression vector of overexpression and knockdown of TFF3 gene were constructed，293T cell was packaged to produce lentiviral particles, virus solution was collected and transfected into TPC-1 cells, enhanced cell TFF3-TPC-1 and enhanced control group ConTFF3-TPC-1; silencing cell shRNA-TFF3-TPC-1 and silencing control cells shCon -TPC-1. Western blotting, and Real-time PCR were used to detect the expression of TFF3 protein and mRNA of four groups. Growth curve and colony formation assay were used to detect the proliferation.Flow cytometry was used to analyze the apoptosis level of the four groups; Western blotting and immunocytochemistry were used to detect apoptosis-related protein and pathway protein phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), nuclear factor-κB（NF-κB）expression.   Results 1. Overexpression and inhibition of expression of TFF3 stable cell TFF3-TPC-1 and shRNA-TFF3-TPC-1 were constructed suscessfully. 2. The proliferation and cloning ability of TFF3-TPC-1 cells were significantly higher than those of ConTFF3-TPC-1 cells（P＜0.05 or P＜0.01）, the proliferation and cloning ability of shRNA-TFF3-TPC-1 cells were significantly lower than those of shCon-TPC-1 cells(P<0.01); 3. The apoptosis rate of TFF3-TPC-1 cells was lower than that of ConTFF3-TPC-1 (0.75%±0.08% vs 5.62%±0.3%, P<0.01),and the apoptosis rate of shRNA-TFF3-TPC-1 was higher than that of shConTPC-1 (22.2% ± 1.2% vs 5.34% ± 0.4%, P<0.01); 4. After silencing TFF3 gene, the expressions of Bax, cytochrome C (Cyt-C), cleavedCaspase-9, cleaved-Caspase-3 were up-regulated, and the expressions of Bcl-2, Akt, p-Akt and NF-κB-P65 were downregulated (P<0.05 or P<0.01).  Conclusion TFF3 may regulate the proliferation and apoptosis of TPC-1 cells by affecting the PI3K/Akt/NF-κB signaling pathway. References | Related Articles | Metrics

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Preparation of honokiol-loaded zein nano-drug delivery system and its anti-breast cancer activity in vitro#br# LIU Dan WANG Jing WANG Liang PENG Yao 2020, 51 (4):  536-542.  doi: 10.16098/j.issn.0529-1356.2020.04.010 Abstract ( )   PDF (6966KB) ( )   Objective To prepare a honokiol (HNK)-loaded zein nano-drug delivery system with low biotoxicity and to investigate its inhibitory effect on MDA-MB-231 breast cancer cells.  Methods Zein-HNK nano-drug delivery system was prepared by anti-solvent method . The particle size, dispersion, morphological characteristics, encapsulation efficiency and in vitro release behavior of the system were investigated. The effects of this system on the proliferation, apoptosis, migration and invasion of MDA-MB-231 cells were evaluated.   Results The particle size of Zein-HNK was (83.75±2.95) nm and the polymer dispersity index(PDI) of it was 0.132±0.015. The encapsulation efficiency of HNK was (93.04±1.86)%. The in vitro release rate of Free-HNK group was(88.90%±2.58)% and release rate of Zein-HNK group was (69.10±1.88）% at 24 hours. Cytotoxicity evaluation showed the 50% inhibiting concentration(IC50) of MDA-MB-231 cells in the positive control group was (5.02±0.29) mg/L, which of MCF-7 cells was (4.72±0.22) mg/L; and the IC50 of MDA-MB-231 cells in Zein-HNK group was (9.92±1.02) mg/L, which of MCF-7 was (9.35±0.30) mg/L. The apoptotic rates of control group, Zein-HNK group and Free-HNK group were (8.73±0.64)%, (15.05±0.85)% and (28.54±1.48)%, respectively. Migration index was 0.67±0.04 in the control group, 0.55±0.03 in the Zein-HNK group and 0.36±0.03 in the Free-HNK group (P<0.05). The relative number of invasions was 0.66±0.03 in the control group, 0.44±0.01 in the Zein-HNK group and 0.35±0.03 in the Free-HNK group (P<0.01). Western bltting result showed that Zein-HNK induced cell apoptosis by decreasing the expression of Bcl-2 and increasing the expression of Bax.   Conclusion Zein-HNK nano-drug delivery system can effectively reduce the toxic and side effects of HNK itself. Zein-HNK nano-drug delivery system can effectively inhibit breast cancer proliferation, induce cell apoptosis, and inhibit cell migration and invasion simultaneously. References | Related Articles | Metrics

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Dauricine inhibiting the cell proliferation and inducing the cell apoptosis of human pancreatic cancer cells line SW1900#br# ZHANG Xiang FAN Jing ZHANG Yi-fen LI Hui LI Chang-yin 2020, 51 (4):  543-547.  doi: 10.16098/j.issn.0529-1356.2020.04.011 Abstract ( )   PDF (1490KB) ( )   Objective To discuss the proliferation inhibition and apoptosis induction of human pancreatic cancer cell line SW1900 by dauricine and its possible mechanism.   Methods The MTT colorimetric method  was used to detect the inhibitory effects of cell viability. The apoptosis rate was tested by the Annexin Ⅴ-FITC/PI fluorescent staining of flow cytometric method . The expressions of phosphatidylinositol 3-kinase (PI3K), protein kinase B(Akt) and B-cell lymphoma-2 (Bcl-2) were detected by Real-time PCR and Western blotting assay.  Results MTT assay showed that dauricine significantly inhibited the proliferation of SW1900 cells and the inhibitory effect was enhanced with the increasing of dauricine concentration, F=783.7, P<0.001. The apoptosis of 3 groups cells were (4.34±1.30)% (0 mg/L dauricine), (14.94±1.94)% (6 mg/L dauricine) and (22.68±3.61)% (12 mg/L dauricine). The mean difference was statistically significant among the three groups (F=58.52, P<0.001). Dauricine could significantly induce apoptosis human pancreatic cancer cells with dose-dependent manner. Real-time PCR showed that the gene expressions of PI3K, Akt and Bcl-2 were lower obviously (PI3K mRNA, F=101, P＝0.01; Akt mRNA, F=1666, P<0.01; Bcl-2 mRNA, F=753, P<0.001) with dose-dependent manner. Western blotting assay also showed that the protein expression of PI3K, Akt and Bcl-2 was down-regulated with dose-dependent manner.  Conclusion Dauricine has proliferation inhibition and apoptosis inducement effect on human pancreatic cancer cells line SW1900. This function may be concerned with the regulation of PI3K/Akt signal pathway and lower Bcl-2 expression.  References | Related Articles | Metrics

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In vitro immune response of insulin-like growth factor 2 mRNA-binding protein 3 restricted cytotoxic T lymphocyte epitope from lung cancer immunotherapy target CHEN Hong-lian SHI Ran-ran ZHANG Li LIU Hui MA Yong-chao 2020, 51 (4):  548-552.  doi: 10.16098/j.issn.0529-1356.2020.04.012 Abstract ( )   PDF (4477KB) ( )   Objective Cancer immunotherapy is attractive for antigen-specific T cell-mediated antitumor therapy, especially in induction of cytotoxic T lymphocytes(CTL). In this report, we evaluated insulin-like growth factor 2 mRNA-binding protein 3(IGF2P3) restricted epitope-induced cytotoxic T lymphocytes effects in human lung cancer cells.   Methods The human leukocyte antigen A2(HLA-A2) restricted epitope peptides of IGF2P3 were selected by NetCTL 1.2, SYFPEITHI and IEDB software prediction. The binding affinity of the peptides to HLA-A02 molecules was evaluated by T2A2 cells binding assay. enzyme-linked immunosport(ELISPOT) assay was used to investigate the ability of the peptide to induce specific restricted cytotoxic T lymphocytes (CTL) and release of interferon γ(IFN-γ). The ability of the peptides to induce T cell response was investigated by carboxyfluorescein succinimidyl amino ester(CFSE) fluorescent staining.   Results The candidate peptide P143, P199, P280, P409 and P515 showed moderate affinity toward HLA-A2 molecule. ELISPOT assay showed P409, P515 were able to induce specific CTL and higher levels of IFN-γ were released. The CTL induced by P409, P515 lysed target cells.  Conclusion P409 and P515 have the potential for adoptive immunotherapy and can be used as candidate epitopes for new anti-tumor polypeptide immunotherapy vaccine. P409 and P515 can be used in peptide-based immunotherapy for lung cancer. References | Related Articles | Metrics

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Expression of miR-145-5p in colorectal cancer and its relationship with multidrug resistance YUAN Ya-nan DU Meng-nan YAN Xiao-li HUANG Yan-jie ZHENG Ji-ning 2020, 51 (4):  553-556.  doi: 10.16098/j.issn.0529-1356.2020.04.013 Abstract ( )   PDF (3209KB) ( )   Objective To investigate the expression and relationship between miR-145-5p and multidrug resistance gene (MDR1) protein P-glycoprotein(P-gp) in colorectal cancer.  Methods SP, Real-time PCR and Western blotting were used to detect the expression of P-gp colorectal carcinoma (CRC) in 50 patients and in 30 paracancerous normal tissue patients, the effect of miR-145-5p expression on MDR1 mRNA and P-gp and the correlation between them and clinicopathological features.   Results The expression level of miR-145-5p in CRC was significantly lower than that in paracancerous normal tissue, and the expression level of MDR1 mRNA and P-gp was significantly higher than that in paracancerous normal tissue (r=-0.403, P<0.01).MiR-145-5p could inhibit the expression of MDR1 mRNA and P-gp in HCT-15 cells(P<0.05).   Conclusion MiR-145-5p plays a regulatory role in the expression of multidrug resistance genes and proteins in colorectal cancer, and is involved in the occurrence, development and multidrug resistance of colorectal cancer. References | Related Articles | Metrics

Anatomy

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High resolution 3D intraosseous vascular model of the scaphoid bone reconstructed by micro CT SHEN Zhong-cheng WANG Ding-yu FANG Xuan LIU Huai-cun JIANG Dong YU Jia-kuo ZHANG Wei-guang 2020, 51 (4):  557-560.  doi: 10.16098/j.issn.0529-1356.2020.04.014 Abstract ( )   PDF (4166KB) ( )    Objective The purpose of this study is to construct a high-resolution model focusing on the vascular pattern of the scaphoid by using micro CT and to provide anatomical reference for the daily clinical use.   Methods The lead-based contrast was perfused from the brachial artery and then the scaphoid bone was harvested. 3D models of the scaphoid bones were constructed by using micro CT to show how arteries distributed in and on the bones.   Results The arteries on the surface stretched from the distal radius covered with scaphoid fossa to the radial side of the waist and then head back to the distal ulna along the dorsoradial ridge, formed like a letter “Ⅴ”. The arteries gathered at the inflection point of the letter “Ⅴ” and the dorsal region. The tubercle region was anastomosed extensively with 3 to 5 major intraosseous vessels originated from the extraosseous vessels covering the waist and the tubercle. There are only 1 to 2 major intraosseous vessels entering the bone via a long route from the ulnar side. The vessels running in the scapholunate ligament didn’t spilt into any intraosseous branches.   Conclusion The superficial vascularity formes a “Ⅴ”-like pattern. The inflection point of the letter “Ⅴ” and the dorsal region display a dense vascularization and these vessels contributed a lot to the intraosseous vascularity. References | Related Articles | Metrics

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Anatomy and clinical significance of the tibiofibular syndesmosis on anticorrosive specimens#br# YANG Jin WANG Ping ZHANG Lei CHENG Han-wen LUO Bei WANG Jun-qiu FENG Qi JIA Wen-li 2020, 51 (4):  561-565.  doi: 10.16098/j.issn.0529-1356.2020.04.015 Abstract ( )   PDF (2782KB) ( )   Objective To research the clinical significance of the tibiofibular syndesmosis based on the anatomical characteristics of the tibiofibular syndesmosis.   Methods A total of 51 tibiofibular syndesmosis collected from Southwest Medical University were measured based on the anatomical characteristics of the lower tibia and fibula of anticorrosive specimens.   Results The geometrical data of the anterior tibiofibular ligament, posterior tibiofibular ligament and transverse tibiofibular ligament were measured respectively, and mean ± standard deviation was described. Include: the length, width, thickness of the anterior tibiofibular ligament (8.53±0.69, 19.06±1.34, 15.99±1.44) mm, the length, width, thickness of the posterior tibiofibular ligament (9.34±0.63, 16.92±1.76, 14.36±0.89) mm, the length, width, thickness of the transverse tibiofibular ligament (18.42±2.48, 21.93±2.59, 4.56±0.17) mm. The angel between the anterior tibiofibular ligament and the coronal surface and the angle of the horizontal plane were (20.49±4.86, 42.20±3.42)°. The angel between the posterior tibiofibular ligament and the coronal surface and the angle of the horizontal plane were (13.2±2.06, 40.92±3.13) °. The angel between the transverse tibiofibular ligament and the coronal surface and the angle of the horizontal plane were (13.45±1.57, 32.73±3.70)°. According to the data analysis, the anterior, posterior and transverse tibiofibular ligaments have statistical difference between men and women, but there is no statistical difference between left and right feet.   Conclusion The tibiofibular syndesmosis is of great significance to the stability of the ankle joint and the anatomical structure has important guiding significance for clinical treatment. References | Related Articles | Metrics

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Anatomy of the safe zone of sustentaculum tali screw fixation LIAO Li-qing CHEN Chen WU Kai LI Yi-kai 2020, 51 (4):  566-569.  doi: 10.16098/j.issn.0529-1356.2020.04.016 Abstract ( )   PDF (3102KB) ( )   Objective To explore the safe zone of sustentaculum tali screw fixation in treatment of calcaneal fracture.   Methods Morphologies of sustentaculum tali in intact dry calcaneus of 589 Chinese adult were investigated and classified. The clinically significant data were observed and measured in sustentaculum tali and calcaneus.   Results Type B calcaneus (77.92%) was defined as the most common type, and type A calcaneus (21.73%) was the second most common type. There were two osseous marks in the medial surface of sustentaculum tali: the posterior tip of middle articular surface for talus and the front end of medial margin of groove for flexor hallucis longus. In the medial of sustentaculum tali, the each side length of quadrilateral was (8.07±1.33)(3.88-12.31)mm, (10.27±1.57)(5.62-15.91) mm, (14.99±2.33) (5.86-21.87) mm and (15.43±2.16) (9.82-22.70) mm, respectively. The length of the its diagonal line was (15.83±2.94)(3.83-26.59) mm. The length of middle articular surface for talus was (18.97±2.85)mm (9.90-25.41).   Conclusion The quadrilateral in the medial surface of sustentaculum tali is the safe zone for sustentaculum tali screw fixation in treatment of calcaneal fracture. References | Related Articles | Metrics

Technology and Methodology

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Establishment of paired box gene 2 knockout mice with CRISPR/Cas9 gene targeting technology WEI Hong-en WANG Min ZHAO Min SHI Xiang-cheng LI Rong-shan 2020, 51 (4):  613-617.  doi: 10.16098/j.issn.0529-1356.2020.04.023 Abstract ( )   PDF (7593KB) ( )   Objective To establish paired box gene 2(Pax2) gene knockout mice by clusterd regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 (Cas9) technology with double nicking method  so as to provide an animal model for studying the role of Pax2 gene in multiple development.  Methods The sgRNA was designed according to the Pax2 gene sequence, and the designed sgRNA and Cas9 were microinjected into the fertilized eggs of C57BL/6J mice in vitro. F0 generation mice were genetically sequenced to identify genotypes after birth, and a total of 8 F0 generation mice were obtained. The F0 mice were mated with wild C57BL/6J mice to obtain F1 mice, and the mice successfully knocked out were successfully mated with C57BL/6J mice to obtain stable Pax2 knockout mice.   Results The stably propagated Pax2 heterozygous knockout mice were successfully obtained, and the Pax2 gene was deleted by 1628 bp. HE staining showed that the number of glomeruli in knockout mice was significantly reduced. Western blotting result  showed that the expression of Pax2 protein in renal cortex of knockout mice was reduced compared with that in wild type mice.  Conclusion The Pax2 heterozygous knockout mice can be successfully constructed by using CRISPR/Cas9 technology, which lays a foundation for further study of the role of Pax2 gene. References | Related Articles | Metrics

Review

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Recent progress of the neural injury mechanism after aneurysmal subarachnoid hemorrhage XIE Jiang-miao YANG Xiao-mei 2020, 51 (4):  618-625.  doi: 10.16098/j.issn.0529-1356.2020.04.024 Abstract ( )   PDF (974KB) ( )   Aneurysmal subarachnoid hemorrhage (aSAH) is one of the most devastating form of stroke. Many physiopathology mechanisms ensue after cerebral aneurysm rupture with blood silting up in the subarachnoid space, including hydrocephalus, cell apoptosis, blood-brain barrier dysfunction, macrovascular vasospasm, microthrombosis and cortical spreading depolarization which interact with each other and work throughout the damage process. Recently, clinical trials gradually pay more attention to two phases of injury after aSAH: the early phase known as early brain injury (EBI) and the delayed phase, delayed cerebral ischemia (DCI). These two phases are main processes accounting for neural injury and are thought to be closely linked with the outcomes of the patients. This review makes a brief summary on the mechanism of cerebral injury after aSAH, mostly on EBI and DCI. References | Related Articles | Metrics

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Research progress on receptor interacting protein kinase regulated necroptosis in nonalcoholic fatty liver disease GUO Yi-ning FAN Qi ZHANG Wei-guang 2020, 51 (4):  626-630.  doi: 10.16098/j.issn.0529-1356.2020.04.025 Abstract ( )   PDF (835KB) ( )   Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease, which might develop into nonalcoholic steatohepatitis (NASH), liver cirrhosis and even carcinoma without effective management. Necroptosis mediated by receptor interacting protein kinase (RIPK) is a novel type of programmed cell death discovered in recent years, which can eventually lead to cell membrane lysis and inflammation. As an important sensor of intracellular and extracellular stress, the RIPK family induces and regulates the activation of necroptosis, and is involved in inflammation and other immune responses. In recent years, studies have shown that RIPK-regulated necroptosis plays an important role in the development of NAFLD. In animal NAFLD/NASH model, the expression of RIPK is related to the degree of hepatic steatosis. In some clinical studies, it was also observed that RIPK expression levels were elevated in NAFLD/NASH patients compared with healthy controls. However, whether necroptosis is a factor which a celerates the progression of liver disease, or a protective factor in the development of liver disease, is still inconclusive. Some studies have shown that RIPK inhibitors may provide guidance for NAFLD treatment. This review provides the molecular mechanisms of necroptosis and its relationship with NAFLD, introduces the important role of RIPK, and summarizes its research progress in the treatment of NAFLD, providing a theoretical basis for further exploration of its mechanism and new treatments for NAFLD. References | Related Articles | Metrics

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Progress of microRNAs in planarian regeneration ZHANG Fen-xi DONG Zi-me, CHEN Guang-wen LIU De-zeng 2020, 51 (4):  631-636.  doi: 10.16098/j.issn.0529-1356.2020.04.026 Abstract ( )   PDF (1389KB) ( )   Planarians have strong regenerative ability, which mainly results from the adult stem cells-neoblasts in their body. Currently, planarians have been an important model organism in the research on regeneration science, and neoblasts have also been the tool cells in biomedical research. But, the scientists still do not find the gene system that effectively regulates planarian regeneration and neoblast proliferation until now. MicroRNAs (miRNAs) are a kind of endogenous, small and noncoding RNAs widely expressed in eukaryotes. MiRNAs can negatively regulate expression of functional genes through specifically recoganizing and binding the target genes. Recently, a few studies have indicated that microRNAs may participate in planarian regeneration. In this review, we will summarize the new progress on miRNAs regulating planarian regeneration as well as neoblast proliferation and differentiation. References | Related Articles | Metrics

Histology,Embryology and Developmental Biology

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Withaferin A protecting against high fat diet induced obesity by inducing white adipose tissue browning GUO Bing-bing WANG Bing-wei SU Zhi-jie ZHANG Chen-yu ZHAO Miao WANG Jun-bo ZHENG Rui-mao 2020, 51 (4):  570-575.  doi: 10.16098/j.issn.0529-1356.2020.04.017 Abstract ( )   PDF (4440KB) ( )    Objective To investigate the protective effect of withaferin A (WA) against high fat diet induced obesity and its associated mechanism.   Methods C57BL/6 J mice at 8-week of age were divided into two groups. The mice were fed with high fat diet (HFD) and were given an intraperitoneal injection of WA or DMSO (solvent control). The body weight and food intake of the mice were monitored. One week later, inguinal white adipose tissue (iWAT), interscapular brown adipose tissue (BAT), epididymal white adipose tissue (eWAT) and retroperitoneal white adipose tissue (rWAT) were collected and weighed. Expression levels of the genes associated with white adipose browning were detected in iWAT. HE staining was applied to observe the morphological changes of iWAT.   Results The data showed that body weight and fat weight in WA group were significantly lower than those in the control group, and the food intake was not changed significantly. Real-time PCR analysis showed that the expression level of browning related genes in iWAT of the WA group was significantly increased. The result  from Western blotting analysis showed that the protein levels of uncoupling protein 1(UCP1) and peroxisome proliferator-activated receptor gamma coactivator 1 alpha(PGC1α) increased significantly in iWAT of the WA group. The typical morphological change of adipose browning, such as the multilocular adipocytes was observed in inguinal white adipose tissue of the  mice treated with WA by using HE staining and mmunofluorescence assay.  Conclusion Taken together, these observations indicate that withaferin A can protect the mice from high fat diet induced obesity by promoting white adipose tissue browning. References | Related Articles | Metrics

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Effects of all-trans retinoic acid on parenchymal organ injury in developmental rats WANG Xia BAI Hao-di SHEN Qin LIU Xing 2020, 51 (4):  576-582.  doi: 10.16098/j.issn.0529-1356.2020.04.018 Abstract ( )   PDF (22016KB) ( )   Objective To investigate the effects of overdose all-trans retinoic acid (ATRA) on brain, heart, lung, liver, kidney and spleen in developmental SD rats.   Methods Forty-eight male SD rats, 3-week-old, were divided into control group and three treatment groups at the doses of 40, 60, and 80 mg/(kg·d) of ATRA, respectively, 12 rats each group, the rats were administrated by gavage every day for 10 days. The rats body weight were weighed every day. After treatment, the rats were sacrificed and then rat heart, lung, liver, kidney, and spleen tissues were weighed, respectively. The organ index was calculated. The organs were then stained with HE staining.   Results After ATRA gavage, compared with the control group, the 40 mg/(kg·d) ATRA group had higher kidney index and body weight change was not statistically significant, the 60 mg/(kg·d) ATRA group had lower body weight, higher heart and kidney index, and lower spleen weight, and the 80 mg/(kg·d) ATRA group had significantly lower body weight, higher brain, heart and kidney index, and lower brain and spleen weight. HE staining showed that compared with the control group, SD rats in the 40, 60 and 80 mg/(kg·d) ATRA groups had thickened alveolar wall, vacuol-like changes in renal tubular epithelial cells, and more macrophages in the spleen, but no significant histological changes in the brain, liver and heart.   Conclusion Excessive all-trans retinoic acid in vivo could damage the lungs, kidneys and spleen in developmental SD rats. References | Related Articles | Metrics

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Effects of peroxiredoxin-1 on pulmonary function in silicotic mice WANG Peng BAI Zhao-rong SHI Qian-wen LI Qian WEI Zhong-qiu LIU Yan AN Hai-juan SUN Ying 2020, 51 (4):  583-587.  doi: 10.16098/j.issn.0529-1356.2020.04.019 Abstract ( )   PDF (5104KB) ( )   Objective To investigate the effect of peroxiredoxin-1 (Prx-1) on pulmonary function induced by silicosis. Methods Forty C57BL/6 mice were randomly divided into control group (normal saline), silicosis group (SiO2), negative control group (SiO2+empty lentivirus) and Prx-1 group (SiO2+Prx1 lentivirus). Crystalline silica instillation and lentivirus were given via trachea, and then all mice were normally raised for 12 weeks. HE staining was used to observe the morphological changes of lung tissue. The expressions of 8-hydroxy-2 deoxyguanosine (8-OHdG), Prx-1, collagen type Ⅰ and Ⅲ were detected by immunohistochemistry and Western blotting respectively. FinePointe WBP whole body plethysmography system was employed to measure the pumonary function.   Results It had the normal structure of lung tissue in control group. SiO2 markedly induced silicosis formation and increase in collagen typeⅠ and Ⅲ, with a similar extent of silicotic nodules and collagen expression in silicosis group and negatively control group, but Prx-1 lentivirus treatment ameliorated these pathological changes. SiO2 increased 8-OHdG and Prx-1 expressions significantly. There were no changes between silicosis group and negative control group. But the level of 8-OHdG in Prx-1 group was lower than silicosis group and Prx-1 level was higher than silicosis group. SiO2 also decreased the pulmonary function significantly. Compared with silicosis group, the mice lung function was improved in Prx-1 group, but there was no effect of empty lentivirus transfection.   Conclusion Overexpression of Prx-1, mediated by decrease in reactive oxygen species(ROS), improves the pulmonary function in mice with silicosis. References | Related Articles | Metrics

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Regular aerobic exercise increasing the expression intestinal mucosal tight junction protein claudin-1 and activating GC-C pathway in facilitating the recovery of obstructive jaundice in mice ZHANG Yu MAO Hai-feng PENG Rui 2020, 51 (4):  588-594.  doi: 10.16098/j.issn.0529-1356.2020.04.020 Abstract ( )   PDF (12575KB) ( )   Objective To investigate the effect of regular aerobic exercise on intestinal mucosal tight junction protein claudin-1 in obstructive jaundice mice and the mechanism of guanylate cyclase C (GC-C) pathway in intestinal mucosal barrier function and inflammatory response.   Methods Thirty male Kunming mice were used to establish the obstructive jaundice model and then were randomly divided into 2 groups: regular aerobic exercise group (EX) and model group (M). In addition, the blank group was set up, 15 mice in each group. ELISA, HE staining, intestinal tissue-associated factor immunohistochemical staining, protein detection, Real-time PCR were used in present study in order to explore the effect of the regular aerobic exercise on the obstructive jaundice of the intestinal mucosal.   Results Yellow discoloration of skin in both abdomen and tail was found in mice in M group. The urine color changed into deep yellow and feces appeared light gray.The bile showed cystic dilatation. Serum total bilirubin (TBIL), total bile acid (TBA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and inflammatory factors interleukin (IL)-8, tumor necrosis factor (TNF)-α protein content significantly elevated. Compared with the M group, the protein content of each indicator in the EX group decreased significantly (P<0.01).Microscopically, the hepatic cord arrangement was disordered in group M, and vacuoles, mitosis, and degeneration appeared in large numbers. The intestinal tissue was sparse and uneven, the intestinal gland and muscle layer disappeared or atrophied, and the interstitial layer was edema and inflammatory cells infiltrated. In the EX group, the intestinal mucosa was thickened, and the number of villi significantly increased compared with the M group and the interstitial showed mild edema. Compared with group M, the positive expression, protein content and gene expression of claudin-1, GC-C, guanylin (Gn), uroguanylin (Ugn) and cyclic guanosine monophosphate (cGMP) in EN group increased significantly (P<0.01).   Conclusion Regular aerobic exercise can effectively promote the expression of tight junction protein Claudin-1 in intestinal mucosa, and activate GC-C pathway to reduce the damages of intestinal mucosal barrier function and inflammatory response.  References | Related Articles | Metrics

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Association between intestinal flora and feces inflammatory factors in recurrent oral ulcer#br# CHEN Xiao-yan ZHU Hong-wei HOU Rui-li LI Zhi-hui LU Jun JIN Mu-lan 2020, 51 (4):  595-604.  doi: 10.16098/j.issn.0529-1356.2020.04.021 Abstract ( )   PDF (12240KB) ( )   Objective To identify whether gut microbial changes were associated with recurrent oral ulcer (ROU) and to determine possible factors associated with the pathogenesis of ROU.  Methods We performed metagenomic sequencing and detected inflammatory cytokines ［interleukin (IL)-10, IL-22］ using ELISA in faeces. Eight students who frequently suffered from oral ulcers were selected as the experimental group (HR); accordingly, 14 healthy volunteers served as the control group (HN).  Results There were some differences in intestinal microbial community structures and core flora between the HR and the HN. At the phylum level, the abundance of Firmicutes in the HR was higher than that in the HN (P<0.05). At the genus level, the abundance of Escherichia/Shigella and Bifidobacterium in the HN were higher than that in the HR (P<0.05). The abundance of enterococcus in the HR was higher than that in the HN (P<0.05). IL-10 levels in feces were similar in HN and HR, and there was no statistical difference between the two groups (P>0.05). IL-22 level in HR was higher than that in HN, and there was statistical significance between the two groups (P<0.05). Univariate Logistic regression analysis showed that IL-22 was a risk factor for ROU. Multivariate Logistic regression analysis showed that Bifidobacterium abundance may be a protective factor for ROU, IL-22 in faeces may be a risk factor. Conclusion Our results suggest that there is an interaction between IL-22 and the distribution of intestinal flora in the patients with ROU, the occurrence of ROU may be related to the decrease of the abundance of Bifidobacteriumand the increase of IL-22 in the faeces, and IL-22 has a stronger proinflammatory effect than the antiinflammatory effect generated by IL-10 and itself in the occurrence of ROU, but further research will be needed. References | Related Articles | Metrics

Neurobiology

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Allopregnanolone restores dopaminergic neurons and motor performance in Parkinson’s disease mice and its molecular mechanisms  WANG Tong-tong CHEN Zhi-chi YE Xin BIAN wei DU Juan-juan FU Wei-da CHEN Meng-jiao LI Jun-nan SUN Chen-you 2020, 51 (4):  473-482.  doi: 10.16098/j.issn.0529-1356.2020.04.001 Abstract ( )   PDF (5918KB) ( )    Objective To investigate the effects of allopregnanolone(APα) on the dopaminergic neurons in substantia nigra, striatal dopaminergic neural fibers and behavioral performance in Parkinson’s disease (PD) model mice, as well as its possible molecular mechanisms.   Methods  C57BL/6 adult male mice with 20-25 g at 3-month old (n=90) were successively injected with 6-hydroxydopamine (6-OHDA) to generate a PD animal model. APα and its receptor γ-aminobutyric acid A receptor (GABAAR) antagonist—bicuculline (Bic) were successively injected. ELISA was used to detect the APα or dopamine concentration in the serum, cerebral cortex and striatum. The number of tyrosine hydroxylase (TH) in the substantia nigra (SN) and striatal dopaminergic neural projections were examined by immunohistochemical staining. The expression levels of GABAAR in membrane fractions and Ca2+/calmodulin-dependent protein kinase Ⅱ δ3 (CaMKⅡδ3), phosphorylated CaMKⅡδ3 (p-CaMKⅡδ3), brain derived neurotrophic factor (BDNF), and cyclin-dependent kinases 1 (CDK1) were detected by Western blotting in the cytoplasmic or nuclear fractions. The interaction of CaMK Ⅱδ3/p-CaMK Ⅱδ3 with BDNF and CDK1 was verified by immunoprecipitation. In addition, the behavioral changes of animals were recorded.   Results There was a significant decrease in the endogenous APα levels of the cerebral cortex, or the dopaminergic neurons of the SN, or the striatal dopaminergic neural fibers, or various protein levels of the membrane, cytoplasmic and nuclear fractions, as well as motor function in PD mice. As compared with PD mice, the abovementioned results were ameliorated in the mice administrated with APα, which were pulled down in mice pre-treated with Bic. The immuno-precipitated result  indicated that CaMKⅡδ3 or p-CaMKⅡδ3 interacted with BDNF or CDK1.   Conclusion  By increasing the endogenous APα levels, exogenous APα treatment can improve SN-striatum dopaminergic neurons in PD model mice by regulating GABAAR/CaMKⅡδ3/BDNF/CDK1 signaling pathway, which in turn promotes behavioral performance in 6-OHDA-lesioned mice. References | Related Articles | Metrics

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Induced experimental autoimmune encephalomyelitis shows the different phenotype of disease in C57BL/6 mice with myelin oligodendrocyte glycoprotein 35-55 MENG Yan-xiao CHU Lan XU Zhu SHAO Bing ZHU Jun-yu Danzhenzhuoma 2020, 51 (4):  483-490.  doi: 10.16098/j.issn.0529-1356.2020.04.002 Abstract ( )   PDF (10272KB) ( )   Objective  To establish an experimental autoimmune encephalomyelitis (EAE)model in female C57BL/6 mice aged 6-8 weeks and investigate its disease phenotype so as to build a better animal model of multiple sclerosis (MS).   Methods  The EAE model was established in 50 female C57BL/6 mice through intradermal injection of myelin oligodendrocyte glycoprotein 35-55 peptide. We assessed the disease progression daily according to a 15-point score for 90 days, and further observed the brain lesions in terms of pathology.   Results  There were three courses of disease in the EAE model: chronic course (3 mice), relapse and remission course (7 mice) and monophasic course (11 mice). Interestingly, we identified some mice (28 mice) had changes in coat color, mental and motor activity, as well as inflammatory demyelinating lesions in the brain tissue although their neurological functions were scored as 0 point.   Conclusion  The EAE model in C57BL/6 female mice shows disease phenotype similar to multiple sclerosis and can be used as a good animal model. References | Related Articles | Metrics

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Changes on dopamin D1 receptor expression in the prefrontal cortex of offspring rats exposed to mobilephone radiation during pregnancy#br# LI Rong WANG Ling-xing LIU Liang-cai ZHANG Yan-qin 2020, 51 (4):  491-494.  doi: 10.16098/j.issn.0529-1356.2020.04.003 Abstract ( )   PDF (3816KB) ( )   Objective To observe the changes of dopamin D1 receptor (D1DR) expression in the prefrontal cortex of offspring rats exposed to mobilephone radiation during pregnancy.   Methods Thirty pregnant rats were randomly divided into three groups namely normal control group, low-dose radiation group and high-dose radiation group.Eighty newborn rats were randomly kept in each group. The offspring rats’prefrontal cortices were collected at day 1, day 10, day 40 and day 60 after birth. The position and intensity of D1DR were detected by immunohistochemistry, and the relative expression of D1DR mRNA was detected by Real-time PCR.   Results The experimental result showed that D1DR was expressed in the cytoplasm, membrane and axon of neurons. The expression intensity of D1DR of the two radiation groups was significantly lower than that of the normal control group(P<0.05) and the expression intensity on D1DR of the high-dose radiation group was significantly lower than that of the low-dose radiation group(P<0.05). The result of Real-time PCR was consistent with those of immunohistochemistry.  Conclusion Exposure to mobile phone radiation during pregnancy could result decreased D1DR expression level in the prefrontal cortex of offspring rats. References | Related Articles | Metrics

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Effect of epigallocatechin gallate on expressions of synaptic plasticity and neural cell adhesion molecules in APP/PS1 double transgenic mice LANG Wei-ya LIU Zhong-jin ZHANG Hai-yan SUN Li-hui ZHU Kun-jie 2020, 51 (4):  495-501.  doi: 10.16098/j.issn.0529-1356.2020.04.004 Abstract ( )   PDF (6431KB) ( )   Objective To observe the effect of epigallocatechin gallate (EGCG) on the spatial learning memory deficit in amyloid procursor protein(APP)/presenilin-1(PS1) double transgenic mice, synaptic ultrastructure and expression of neural cell adhesion molecule in hippocampal CA1 region.   Methods Eight weeks old male APP/PS1 double transgenic mice were selected as Alzheimer’s disease (AD) model and divided into the model group, the EGCG group and the donepezil hydrochloride group, 12 in each group.Besides，normal mice of the same brood (with no transgene) were recruited as a normal group (n=12). Related indices were detected after 6 months continuous gastrogavage. The spatial learning-memory deficit of APP/PS1 double transgenic mice was detected by Morris water maze test.The synaptic ultrastructure of hippocampal CA1 region was observed by transmission electron microscopy. The expression levels of neural cell adhesion molecule (NCAM) and polysialyltranseferase α2,8-polysialic acid (ST8Sia Ⅱ) protein in hippocampal CA1 region of APP/PS1 transgenic mice were detected by immunofluorescence and Western blotting.   Results Compared with the normal group, the mean value of escape latency in the model group was extended, and compared with the model group, the mean value of escape latency in the EGCG group and donepezil hydrochloride group were increased (P<0.05). The result  of electron microscope showed that the changes of synaptic interface curvature of EGCG group and donepezil hydrochloride group were not obvious. Compared with the model group, the width of the synaptic gap becomes narrower and the thickness of the postsynaptic compact were increases (P<0.05). Immunofluorescence showed that the expression of NCAM and ST8Sia Ⅱ proteins in the hippocampus CA1 region was expressed in the cytoplasm of neurons, the expressions of NCAM and ST8Sia Ⅱ in hippocampal CA1 region were significantly increased in EGCG group and donepezil hydrochloride group (P<0.05). Their contents also showed higher levels of expression in Western blotting (P<0.05).  Conclusion EGCG shows improvement on the spatial learning-memory deficit in APP/PS1 double transgenic mice，which may be associated with affecting the synaptic structure of hippocampus and improving the expressions of neural cell adhesion molecule. References | Related Articles | Metrics

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Effect of spinal cord decellularized scaffold on the repair of spinal cord defects in rats#br# ZHU Jun-yi ZHANG Yi-jia DU Sheng-hu YU Fang-zheng LU Ying-feng LI Pei-feng WANG Zhi-bin MEI Jin WANG Jian 2020, 51 (4):  502-506.  doi: 10.16098/j.issn.0529-1356.2020.04.005 Abstract ( )   PDF (5529KB) ( )   Objective Exploring the effect of spinal cord decellularized scaffold on spinal cord defects and observing the behavior and regeneration of rats after operation.   Methods The spinal cords of 30 SD rats were treated with 3% Triton X-100 and 2% sodium deoxycholate on oscillator. The cell residue and the spatial structure of the tissue were compared before and after treatment, in order to understand the tissue structure of the stent itself. 90 SD rats were randomly divided into control group, simple injury group and stent transplantation group. Excision of the spinal cord 9-10 segments in the simple injury group and the stent graft group the acellular scaffold was transplanted to the stent graft group. Behavioral scores were observed postoperatively. At 4, 8, and 12 weeks, the spinal cords of the injured part of the rats were taken for HE staining and immunofluorescence detection of nerve regenerationrelated proteins.  Results After decellularization of the spinal cord, the nerve cells and axons were completely removed, and the extracellular matrix of the spinal cord was preserved. Scanning electron microscopy revealed that the scaffold retained a certain porous network scaffold structure. In the experiment of decellularized scaffold in vivo, the Basso-Beattie-Bresnahan(BBB) score showed that the recovery of hindlimb motor function in rats with decellularized scaffolds was better than that in rats with simple injury. HE staining showed that the decellularized scaffold could fill the defect of the spinal cord segment and accelerate the repair process of the injured spinal cord. Immunofluorescence showed that there was a certain axonal regeneration in the injured part of the stent transplantation group.  Conclusion The spinal cord decellularized scaffold retains the extracellular matrix and has a certain spatial structure, which can accelerate the process of spinal cord defect repair to a certain extent, and has a certain promoting effect on nerve regeneration. References | Related Articles | Metrics

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Detection of intestinal microbial changes in rotenone-induced Parkinson’s disease model mice#br# HAN Qiu-qin WAN Guo-qing GU Xue-feng 2020, 51 (4):  507-512.  doi: 10.16098/j.issn.0529-1356.2020.04.006 Abstract ( )   PDF (2482KB) ( )   Objective To investigate the changes of intestinal microbes in rotenone-induced Parkinson’s disease (PD) mice based on 16S rRNA gene sequencing.   Methods Fourteen 8-weekold male C57BL/6J mice were randomly divided into two groups: 6 mice in the control group and 8 mice in the model group. The model mice were injected subcutaneously with rotenone (3 mg/kg) for 5 weeks, and the body weight was measured once a week. After 5 weeks, behavioral tests were performed, including the rotating rod test and the open field test. The contents of the tract were used for intestinal microbial detection analysis.  Results After 5 weeks of rotenone treatment, the weight of PD mice was significantly lower than that of the control mice（P<0.05）. The movement time of the PD model mice in the rotating rod（P<0.0001）and the total distance of movement in the open field（P<0.05）were significantly lower than that of the control mice. In addition, the intestinal microbial diversity of the PD model mice did not change（P>0.05）,but the microbial species showed significant differences. Among them, the PD mice showed a significant decrease in the intestinal  Turicibacter（P<0.01）, a significant increase in norank_f_Lachnospiraceae（P<0.01）, a significant decrease in norank_f_ Erysipelotrichacea e（P<0.01）, and a significant increase in Lachnoclostridium（P<0.01）.  Conclusion Intestinal microbes in PD mice are disordered, and these intestinal flora may be involved in the development of dyskinesia in PD mice. References | Related Articles | Metrics

Cell and Molecules Biology

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Experiment of the differentiation of adipose-derived stem cells into fibroblasts JU Xiao-jun NIU You-ya RAO Li-bing YI Li-ming XIANG Sha ZHANG Ying-jun 2020, 51 (4):  513-519.  doi: 10.16098/j.issn.0529-1356.2020.04.007 Abstract ( )   PDF (14184KB) ( )   Objective To explore the possibility of adipose-derived stem cells（ADSCs）to differentiate into fibroblasts, and to provide an effective way for the effective solution of skin tissue engineering seed cells.   Methods Primary ADSCs were obtained from inguinal fat of ten healthy adult SD rats，weighing 280-320 g，and cultured in vitro and purified.When primary ADSCs expansion to the 3rd passages，the following experiments were performed alkaline phosphatase test on the 16th day after osteogenesis induction，staining of alizarin red mineralized nodules on day 23 after osteogenesis induction，oil red O staining on day 12 after adipogenic induction；Flow cytometry detection of cell surface markers；Addition of conditioned medium to induce differentiation into fibroblasts，Photograph the changes of cell morphology on the 2nd, 4th, 6th, and 8th days after induction，MTT to detect cell viability at various time points；Scanning electron microscopy on day 6 and day 8 after induction；Immunocytochemical staining on day 8 after induction，detect the expression of vimentin, the main marker of fibroblasts.  Results Primary ADSCs grew in long spindles，showed strong positive expression of alkaline phosphatase (ALP) after osteogenesis induction，and alizarin red staining showed red mineralized nodules；Aggregation of intracellular red-stained lipid droplets after adipogenic induction were found；Flow cytometry showed positive expression of mesenchymal stem cell-related marker CD90，and hematopoietic stem cell marker CD45 was negative. Morphology of ADSCs started to change on day 2 after induction into fibroblasts. On the 4th day after induction, the cells were in the shape of water droplets or short rods. On the 6th day after induction, the cells were protruded polygonal or triangular. Cells crowded and covered the bottom of the bottle on day 8 after induction，becoming slender fibrous. MTT test showed that the cell viability was significantly lower on the second day after induction than in the control group. There were no significant differences in cell viability on the 4th, 6th, and 8th days after induction compared with the control group. Scanning electron microscopy showed that the cells were triangular on the 6th day after induction, and the surface had more cilia. On the 8th day after induction, the cells were slender and fibrous, with small protrusions, and the surface cilia were dense. Vimentin was positively expressed in most cells on the 8th day after induction.  Conclusion ADSCs can have the morphological characteristics of fibroblasts after induced differentiation in vitro；that can express fibroblasts marker protein. References | Related Articles | Metrics

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Role of fibroblast growth factor 2 and tanshinoneⅡA in the differentiation of rat bone marrow mesenchymal stem cells into cardiomyocyte-like cells LI Jiao Lü Yang LIU Yang WANG Hao-yu ZHAO Ya-ru WANG Xue-jiao WANG Hai-ping 2020, 51 (4):  520-527.  doi: 10.16098/j.issn.0529-1356.2020.04.008 Abstract ( )   PDF (10011KB) ( )   Objective To investigate the effects of fibroblast growth factor 2 (FGF-2) combined with tanshinoneⅡA on differentiation of rat bone marrow mesenchymal stem cells (BMSCs）into cardiomyocyte-like cells.  Methods BMSCs were isolated and cultured. The cultured cells were identified by flow cytometry, BMSCs were divided into experimental control group, FGF-2 group, tanshinoneⅡA group and the combined induction group. The activity and value of  BMSCs were detected by MTT. The Real-time PCR method was used to detect BMSCs. Expression of early myocardial transcription factors GATA-4 and Nkx2.5; Immunocytochemical staining for detection of connexin43（Cx43）and cardiac troponin-Ⅰ(cTnI);Immunofluorescence staining for detection of desmin and Tm; The expression of desmin and tropomyosin（Tm）was detected by Western blotting.  Results The cell activity and proliferation after induction were good. The expression of GATA-4 and Nkx2.5 in the induction group were stronger than that in the experimental control group, and the difference was statistically significant (P<0.05). The positive expression of Cx43 and cTnI in the induction group increased significantly. The markers of the combined group were the most obvious, and the difference was statistically significant (P<0.05). The expression of Myo-specific protein desmin and Tm in the combined induction group increased significantly, and the difference was statistically significant (P<0.05).  Conclusion Both FGF-2 and tanshinoneⅡA can promote the proliferation of BMSCs and induce the differentiation of BMSCs into cardiomyocyte-like cells. The synergistic effect of the two is better than other groups. References | Related Articles | Metrics

Anthropology

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Regional differences of nasal discrete traits in Han Chinese LI Zi-jun WEI Pian-pian SUN Chang WEN Shao-qing TAN Jing-ze 2020, 51 (4):  605-612.  doi: 10.16098/j.issn.0529-1356.2020.04.022 Abstract ( )   PDF (2170KB) ( )   Objective To explore the gender and regional differences of the nasal discrete traits in Han ethnic groups.   Methods From 2015 to 2019, eight nasal discrete traits were observed in three Han ethnic groups from Henan, Jiangsu and Guangxi, with sample size of 1022, 963, and 1017, respectively. The statistical tests were performed on gender and regional level, and the multiple correspondence analysis (MCA) was used to study the correlation between these morphological traits and regions.  Results Most nasal discrete traits showed significant statistical differences within gender, and exhibit differences in regional populations. In dimension 1 (Dim 1) of MCA figures, samples in Guangxi separate from the other two regions’, while there was considerable overlap between samples from Henan and Jiangsu. Among eight nasal discrete traits, the shape of nasal tip, alae nasi height and alae nasi projection contribute most to Dim1.  Conclusion Nasal tip and nasal wings exhibited significant regional differences in Han Chinese. References | Related Articles | Metrics