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Journal of Jilin University(Medicine Edition)
ISSN 1671-587X
CN 22-1342/R
主 任:王 丽
编 辑:姜瑾秋 李欣欣 韩宏志
    官 鑫
电 话:0431-85619279
E-mail:xuebao@jlu.edu.cn
地 址:长春市新民大街828号
    (130021)
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Current Issue
28 July 2020, Volume 46 Issue 04
Research in basic medicine
Effects of down-regulation of ADAR1 expression on proliferation, migration and epithelial-mesenchymal transition of human lung cancer cells
ZHOU Ning, WU Rui, MA Zhenkai, CHEN Weiwei, LI Xuelin, GONG Kaikai, YANG Lijuan, DAI Juanjuan, WU Yan
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  669-674.  DOI: 10.13481/j.1671-587x.20200401
Abstract ( 40 )   PDF (662KB) ( 36 )  
Objective: To investigate the effects of down-regulation of adenosine deaminase acting on RNA enzyme 1 (ADAR1) on the proliferation, migration and epithelial-mesenchymal transition (EMT) of the lung cancer H1299 and H520 cells, and to elucidate their mechanisms. Methods: The H1299 and H520 cells were divided into control group (transfected with negative shRNA) and shADAR1 group (transfected with shADAR1). Real-time fluorescence quantitative PCR and Western blotting methods were used to detect the expression levels of ADAR1 mRNA and protein, respectively. CCK8 method and colony information assay were used to detect the proliferation activities and the amounts of colony formation;scratch assay was used to detect the scratch healing rates of the cells in various groups; Western blotting method was performed to detect the expression levels of E-cadherin and vimentin proteins in the cells in various groups. Results: Compared with control group, the expression levels of ADAR1 mRNA and protein in the H1299 and H520 cells in shADAR1 group were significantly decreased(P<0.05).Compared with control group, the proliferation activity, the colony formation amount and the scratch healing rate of the H1299 and H520 cells in shADAR1 group were significantly decreased (P<0.05).Compared with control group, the expression levels of E-cadherin protein in the H1299 and H520 cells in shADAR1 group were significantly increased(P<0.01), and the expression levels of vimentin protein were decreased (P<0.01). Conclusion: Down-regulation of ADAR1 expression contributes to inhibiting the proliferation, migration, and EMT of lung cancer cells.
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Dynamic expression and localization of CDC14B in each cell cycle of mouse one-cell fertilized eggs
LIU Lanqing, LI Ruilin, ZHOU Xiaomin, MENG Jun
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  675-679.  DOI: 10.13481/j.1671-587x.20200402
Abstract ( 11 )   PDF (484KB) ( 12 )  
Objective: To observe the expression and localization of cell division cycle 14B (CDC14B) in each cell cycle of the mouse one-cell fertilized eggs,and to elucidate preliminarily the mechanism of CDC14B regulating the early development of mammalian fertilized eggs. Methods: The superovulation technique was used to collect the mouse one-cell fertilized eggs. The expression levels of CDC14B mRNA and protein in the mouse one-cell stage fertilized eggs in different cell cycles were detected by RT-PCR and Western blotting methods, respectively. Co-localization of CDC14B and β-tubulin in the mouse one-cell fertilized eggs in different cell cycles was observed by cell immunofluorescence method. Results: Compared with G1 phase, the expression levels of CDC14B mRNA and protein in the mouse one-cell fertilized eggs in S, G2 and M phases were significantly increased(P<0.05 or P<0.01).The cellular immunofluorescence assay results showed that CDC14B (red fluorescence) and β-tubulin (green fluorescence) were co-located in the cortex of fertilized mouse eggs in G1 phase and S phase,in the early G2 phase, CDC14B and β-tubulin co-localized in the cortex and cytoplasm; in the late G2 phase, part of CDC14B entered the nucleus; in the M phase, CDC14B and β-tubulin co-localized in the spindle. Conclusion: CDC14B dynamically expresses in each cell cycle of the mouse one-cell fertilized eggs. CDC14B has nuclear plasma spindle translocation, and the localization changes of CDC14B may regulate the spindle function.
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Inhibitory effect of Lactobacillus rhamnosus on intestinal inflammation after spinal cord injury in zebrafishes and its mechanism
ZHAO Liping, HUANG Shubing, ZHANG Boping, ZHOU Zhilan, JIA Xuebing, SUN Mengfei, QIAO Chenmeng, CHEN Xue, SHEN Yanqin, CUI Chun
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  680-686.  DOI: 10.13481/j.1671-587x.20200403
Abstract ( 8 )   PDF (875KB) ( 6 )  
Objective: To study the influence of spinal cord injury (SCI) in intestinal inflammation in the zebrafishes, and to clarify the effect of alleviating the intestinal inflammation of Lactobacillus rhamnosus GG (LGG) and its mechanism. Methods: Fourteen normal zebrafishes were randomly divided into normal diet group (n=7) and LGG group (n=7).After 21 d of feeding, intestinal tissues were taken and the expression levels of intestinal barrier-related molecules defensin beta-like 1 (DEFBL1), tight junction protein 2a (TJP2a), mucin 2.1 (MUC2.1) and mucin 5.3 (MUC5.3) in intestinal tissue of the zebrafishes were detected by RT-qPCR method. Forty eight operated zebrafishes were randomly divided into sham operation + normal diet group (n=12), sham operation + LGG group (n=12), SCI + normal diet group (n=12) and SCI + LGG group (n=12).After 25 d of feeding after injury (n=6 in each group), intestinal tissues were taken and the expression levels of intestinal inflammatory cytokines tumor necrosis factor-α (TNF-α), Toll-like receptor 2 (TLR-2) and interleukin-6 (IL-6) were detected by RT-qPCR method. After 7 d (n=3 in each group) and 28 d (n=3 in each group) of feeding after injury, the intestinal tissues were taken for high-throughput 16S rRNA sequencing of intestinal flora to detect the changes in intestinal flora. Results: Compared with normal diet group,the expression levels of DEFBL1 and TJP2a in intestinal tissue of the zebrafishes in LGG group were significantly increased(t=-2.387, P<0.05;t=-12.482, P<0.001), but the expression levels of MUC2.1 and MUC5.3 were significantly decreased (t=2.497, P<0.05;t=3.173, P<0.05). Compared with sham operation +normal diet group, the expression levels of TNF-α, TLR-2 and IL-6 mRNA in the intestinal tissue of the zebrafishes in SCI + normal diet group were increased significantly(P<0.05 or P<0.01);compared with SCI + normal diet group, the expression levels of TNF-α, TLR-2, and IL-6 in intestinal tissue of the zebrafishes in SCI + LGG group were decreased(P<0.05 or P<0.01).Compared with normal diet groups (sham + normal diet group and SCI + normal diet group), the abundance of pathogenic bacteria (Aeromonas and Vibrio) and conditioned pathogenic bacteria (Desulphomonas and Brevundimonas) in intestinal tissue of the zebrafishes in sham + LGG group and SCI + LGG group were significantly decreased. Conclusion: LGG can inhibit the intestinal inflammation caused by SCI in the zebrafishes, and its mechanism may be related to increasing the expression levels of intestinal barrier-related molecules and decreasing the abundance of intestinal pathogenic bacteria.
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Fluorescence labelingand identification of panax ginseng active glycopeptide and its distribution in main organ tissues of mice
FANG Xiaoxue, XU Wei, ZHOU Tingting, TANG Yan, WANG Chunchi, JIANG Ruizhi, QIU Zhidong, LUO Haoming
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  687-692.  DOI: 10.13481/j.1671-587x.20200404
Abstract ( 6 )   PDF (910KB) ( 7 )  
Objective: To observe the distribution of panax ginseng active glycopeptide (PGG) in the main organ tissues of the mice and its targeting trend by fluorescence labeling. Methods: PGG was labeled with fluorescein isothiocyanate (FITC) as a probe. The structures of PGG fluorescence marker (PGG-FITC) were analyzed by ultraviolet spectrophotometry and infrared spectrophotometry. The fluorescence substitution degree of PGG-FITC was determined by fluorescence spectrophotometry. A total of 78 mice were randomly divided into 13 groups; the mice in 6 groups were injected with PGG-FITC, the mice in another 6 groups were injected with FITC, and the residual group was used as blank control. The heart, liver, spleen, lung, kidney, brain, intestine and stomach tissues of the mice in various groups were collected at 0.5, 1.0, 2.0, 4.0, 8.0 and 24.0 h after administration. The levels of PGG-FITC and FITC in the main organ tissues of the mice were determined by fluorescence spectrophotometry, and the area under concentration time curve (AUC) and targeting efficiency (Te) were calculated. Results: PGG and FITC were linked by covalent bond and the fluorescence substitution degree was 1.435%. Compared with FITC group, the drug levels in main organ tissues of the mice in PGG-FITC group were increased significantly (P<0.01), and the drug levels in brain tissue were increased more significantly. The AUC values in main organ tissues of the mice in FITC group from high to low were liver, brain, stomach, kidney, intestine, heart, spleen and lung tissues;and the AUC values in main organ tissues of the mice in PGG-FITC group from high to low were brain, heart, liver, kidney, spleen, stomach, lung and intestine tissues. Conclusion: PGG labeled by FITC is mainly enrich in the brain tissue of the mice after tail vein injection of PGG, and has a brain targeting trend.
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Enhancement of mitochondria-targeted KillerRed in autophagy caused by radiation in HeLa cells and its mechanism
YU Lei, WANG Ce, HAN Bing, LI Xin, HAN Yuchen, SUN Yuying, GUO Xiangshu, LIU Weiwu, WANG Zhicheng
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  693-698.  DOI: 10.13481/j.1671-587x.20200405
Abstract ( 7 )   PDF (869KB) ( 7 )  
Objective: To explore the enhancement of mitochondira-targeted KillerRed(mtKR) on the mitochondrial dysfunciton and autophagy caused by radiation in the HeLa cells, and to clarify the relative molecular mechanisms. Methods: After mitochondria-targeted expression vectors Pink-mtKR were transfected into the HeLa cells, the cells were irradiated by visible light and 4 Gy X-ray. The cells were divided into control, empty vector, Pink1-mtKR, control + 4 Gy X-ray irradiation, empty vector + 4 Gy X-ray irradiation and Pink1-mtKR+4 Gy X-ray irradiation groups. After the cells were irradiated with X-ray for 24 h, the mitochondrial membrane potentials (MMP) and the autophgic rates were detected by flow cytometry, the levels of mitochondrial respiratory chain complex Ⅰ and Ⅲ were measured by biochemical assay,and the expression amounts of P62, Pink1, parkin and Tom20 proteins were measured by Western blotting method. Results: After Pink1-mtKR plasmids were transfected into the HeLa cells, the cells were irradiated by visible light and 4 Gy X-ray; compared with control group, the MMP and the levels of mitochondrial respiratory chain complex Ⅰand Ⅲ in Pink1-mtKR group were significantly decreased (P<0.05), and the autohpagic rate was significantly increased (P<0.05); the MMP and the levels of mitochondrial respiratory chain complex Ⅰ and Ⅲ in Pink1-mtKR + 4 Gy X-ray irradiation group were decreased obviously (P<0.05), and the autohpagic rate was significantly increased(P<0.05). Compared with control group, the expression amounts of Pink1 and parkin proteins in total protein of the cells in Pink1-mtKR group and Pink1-mtKR+4 Gy X-ray irradiation group had no obvious changes, but the expression amounts of the P62 protein was increased; the expression amounts of Tom20, Pink1 and parkin in mitochondrial protein in Pink1-mtKR group and Pink1-mtKR+4 Gy X-ray irradiation group were increased obviously. Conclusion: MtKR may enhance the mitochondrial dysfunction and autohpagy caused by radiation, and its mechamism may be related to the regulation of Pink1/parkin autophgy pathway.
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Effect of oncostatin M on proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells
KONG Ning, ZHOU Yulai, ZHANG LU, SHI Yi, SHI Yan, SUN Zhongping, ZOU Yinggang
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  699-706.  DOI: 10.13481/j.1671-587x.20200406
Abstract ( 11 )   PDF (1184KB) ( 7 )  
Objective: To investigate the effect of oncostatin M (OSM) on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells(hUCMSCs), and to elucidate that OSM is an effective osteogenic induction active factor. Methods: The hUCMSCs were cultivated in vitro. The surface antigens of hUCMSCs at the third generation were detected by flow cytometry. CCK-8 assay was used to determine the proliferation activities of hUCMSCs after treated by 0.1,1.0 and 10.0 μg·L-1 recombined human oncostatin M(rhOSM).The hUCMSCs were divided into control group,classical osteogenic induction group,and different doses (0.1,1.0 and 10.0 μg·L-1) of rhOSM osteogenic induction groups. Alkaline phosphatase(ALP) was stained by BCIP/NBT and the ALP activities in the cells were determined quantitatively on the 4th,7th and 14th days of osteogenic induction; Alizarin Red S staining (ARS) was performed to assess the formation of calcium nodules and the cell mineralization activities were semi-quantitatively analyzed. RT-qPCR was carried out to measure the expression levels of Runt-related transcription factor 2(Runx2) mRNA and OCN mRNA in the hUCMSCs. Results: The hUCMSCs were consistent with the identification criteria of mesenchymal stem cells(MSCs).After treatment with rhOSM for 72 h, compared with control group, the proliferation activity of hUCMSCs in 10.0 μg·L-1 rhOSM group was decreased (P<0.05); after treated for 96 h, the proliferation activities in different doses of rhOSM groups were significantly lower than that in control group (P<0.05); there were significant differences between different doses of rhOSM groups(P<0.01).On the 4th, 7th and 14th days,compared with classical osteogenic induction group,the ALP activities and the expression levels of Runx2 mRNA in the hUCMSCs in different doses of rhOSM groups were increased(P<0.01).Compared with 0.1 μg·L-1 rhOSM group,the ALP activities and the expression levels of Runx2 mRNA in the hUCMSCs in 1.0 and 10.0 μg·L-1 rhOSM groups were significantly increased(P<0.05). Compared with the 7th day of induction,the ALP activities and the expression levels of Runx2 mRNA in different doses of rhOSM groups were slightly decreased on the 14th day of induction, but there were no significant differences(P<0.05).On the 7th,14th and 21st days,compared with classical osteogenic induction group, the mineralization activities in the hUCMSCs in different doses of rhOSM groups were increased(P<0.01);compared with 0.1 μg·L-1 rhOSM group,the mineralization activities in the hUCMSCs in 1.0 and 10.0 μg·L-1 rhOSM groups were significantly increased(P<0.05).Compared with classical osteogenic induction group,the expression levels of OCN mRNA ih the hUCMSCs in different doses of rhOSM groups were significatnly increased(P<0.05);on the 7th,14th and 21st days of induction,compared with 0.1 μg·L-1 rhOSM grouop,the expression levels of OCN mRNA in the cells in 1.0 and 10.0 μg·L-1 rhOSM groups were signficantly increased(P<0.05) in a time-dose manner. Conclusion: rhOSM can promote the osteogenic differentiation of hUCMSCs in vitro by increasing the expressions of Runx2 and OCN and increasing the ALP activity and the deposition of calcium salts and is an effective osteogenic induction active factor.
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Improvement effect of ginsenoside Rg1 combined with ginsenoside Rg3 on reproductive function in male model mice with reproductive function injury
LI Huan, QU Zhenting, QIAN Honghao, XU Han, QU Li, ZHANG Jing, WANG Hongyan, XU Xiaolei
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  707-713.  DOI: 10.13481/j.1671-587x.20200407
Abstract ( 13 )   PDF (1222KB) ( 11 )  
Objective: To investigate the improvement effect of ginsenoside Rg1 combined with ginsenoside Rg3 on the reproductive function in the dinbutyl phthalate (DBP)-induced reproductive function injury model mice, to elucidate the mechanism of ginsenoside Rg1 combined with ginsenoside Rg3 in the protection of reproductive function, and to provide the experimental basis for their further development. Methods: Twenty-eight clean grade male C57BL/6 mice were randomly divided into model group (400 mg·kg-1 DBP),ginsenoside Rg1+DBP group (20 mg·kg-1 ginsenoside Rg1+400 mg·kg-1 DBP),ginsenoside Rg3+DBP group (20 mg·kg-1 ginsenoside Rg3+ 400 mg·kg-1 DBP) and combination group (20 mg·kg-1 ginsenoside Rg1+20 mg·kg-1ginsenoside Rg3+400 mg·kg-1 DBP); there were 7 mice in each group. DBP dissolved in corn oil was given to the mice by gavage every morning,ginsenoside Rg1 and ginsenoside Rg3 were dissolved in normal saline and injected subcutaneously in the mice every afternoon, and DBP,ginsenoside Rg1 and ginsenoside Rg3 were administered once a day for 35 consecutive days. The sperm density, sperm viability and total malformation rate of the mice in each group were detected by Wljy-9000 sperm quality detection system. HE staining was used to observe the patholmorphology of testis tissue of the mice. The levels of serum testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were detected by enzyme-linked immunosorbent assay (ELISA). The expression level of Cx43 protein in testis tissue was detected by immunofluorescence assay. Results: Compared with model group, the testicular organ coefficient of the mice in combination group was significantly increased (P<0.05), and the sperm densities and the sperm viabilities of the mice in ginsenoside Rg1+DBP group, ginsenoside Rg3+DBP group and combination group were significantly increased (P<0.01). Compared with model group, the levels of serum T and LH of the mice ginsenoside Rg1+DBP group, ginsenoside Rg3+DBP group and combination group were significantly increased(P<0.01) and the FSH levels were significantly decreased in(P<0.01); the expression levels of Cx43 protein were significantly increased (P<0.05). The HE staining results showed that the spermatogenic epithelium of the testis tissue of the mice in model group was relatively thin, and the spermatogenic cells in the duct cavity were seriously exfoliated; compared with model group, the spermatogenic epithelium of the testis tissue of the mice in ginsenoside Rg1+DBP group and ginsenoside Rg3+DBP group became thicker, and fewer exfoliated cells were found in the lumen; the spermatogenic epithelial structure of the testis tissue in combination group was complete, with regular spermatogenic cells at all levels and abundant intracellular sperm. The result of factorial analysis showed that there was an interaction between ginsenoside Rg1 and ginsenoside Rg3, which resulted in the increase of sperm viability(F=6.704, P=0.016), the serum T level(F=4.912, P=0.036)and the expression level of Cx43 protein in testis tissue(F=6.937, P=0.018). Conclusion: Ginsenoside Rg1 and ginsenoside Rg3 used alone or combined can improve the sperm density and the sperm viability, increase the serum T and LH levels, decrease the FSH level in the mice, and reduce the reproductive function injury in the mice, and its mechanism might be related to the protective role in spermatogenesis by up-regulating the expression level of Cx43.Ginsenoside Rg1 combined with ginsenoside Rg3 has a synergistic effect.
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Sedative and hypnotic effects of water and alcohol extracts from fruits of Acanthopanax senticosus in mice and their possible mechanisms
WANG Jingyao, ZHAO Yan, CAI Enbo, ZHU Hongyan, LI Pingya, LIU Jinping
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  714-721.  DOI: 10.13481/j.1671-587x.20200408
Abstract ( 5 )   PDF (1411KB) ( 5 )  
Objective: To explore the sedative and hypnotic effects of Acanthopanax senticosus fruit water extract (ASF-WE) and alcohol extract (ASF-AE) in the mice, to elucidate the possible mechanisms, and to provide the basis for the development of Acanthopanax senticosus. Methods: ASF-WE and ASF-AE were prepared, respectively. A total of 120 male ICR mice were randomly divided into blank control group, diazepam(DZP) positive control group(DZP group), different doses (4, 8, 16, 32 and 64 mg·kg-1) of ASF-WE groups, and different doses (4, 8, 16, 32 and 64 mg·kg-1) of ASF-AE groups, 10 mice in each group, continuous administration for 5 d. The number of locomotor activity of the mice in various groups was recorded. The sleep rates of mice were recorded by using subthreshold dose of pentobarbital sodium induced sleep experiment. The sleep latency and sleep time of mice were recorded by using threshold dose of pentobarbital sodium induced sleep experiment. The hypnotic dose and subhypnotic dose of ASF-WE and ASF-AE were selected. According to the different model drugs, the model drugs were 5-hydroxytryptophan (5-HTP), flumazenil (FLU) and p-corophenylalanine (pCPA),40 male ICR mice were randomly divided into blank control group, model control group(5-HTP group and pCPA group), ASF-WE + model drug group and ASF-AE + model drug group, 10 mice in each group;80 male ICR mice were randomly divided into blank control group,FLU group,DZP group,ASF-WE group,ASF-AE group,DZP+FLU group,ASF-WE group and ASF-AE+FLU group,10 mice in each group, for 5 consecutive days. Pentobarbital sodium induced sleep experiment was used to record the sleep latency and sleep time of mice. ELISA was used todetect the levels of 5-hydroxytryptamine(5-HT) and gamma-aminobutyric acid (GABA) in hippocampus tissue of the mice. Results: Compared with control group, the number of locomotor activity of the mice in 32 and 64 mg·kg-1 ASF-WE groups were significantly decreased (P<0.05 or P<0.01),and the number of locomotor activity of the mice in 16,32 and 64 mg·kg-1ASF-AE groups were significantly decreased (P<0.05 or P<0.01). Compared with blank control group, the sleep latencies of the mice in 16,32 and 64 mg·kg-1 ASF-WE groups were significantly reduced(P<0.05 or P<0.01),and the sleep time was significantly prolonged (P<0.05 or P<0.01); the sleep latencies of the mice in 16,32 and 64 mg·kg-1 ASF-AE groups were significantly shortened(P<0.05 or P<0.01), and the sleep time was significantly prolonged (P<0.05 or P<0.01).Compared with 5-HTP group, the sleep latencies of the mice in ASF-WE + 5-HTP group and ASF-AE + 5-HTP group were significantly shortened (P<0.01), the sleep time was significantly prolonged (P<0.01),and the levels of 5-HT in rat hippocampus tissue were significantly increased (P<0.05). Compared with blank control group, the sleep latency of the mice in pCPA group was significantly prolonged (P<0.01), the sleep time of the mice was shortened (P<0.01), indicating that the insomnia model was successfully established. Compared with pCPA group, the sleep latencies of the mice in ASF-WE + pCPA group and ASF-AE + pCPA group were significantly shortened (P<0.05),the sleep time was significantly prolonged (P<0.01), and the GABA levels in hippocampus tissue of the mice were significantly increased (P<0.05 or P<0.01). Compared with ASF-WE group, the sleep latency of the mice in ASF-WE+FLU group was significantly prolonged (P<0.05), the sleep time was significantly shortened(P<0.01), and the GABA level in the hippocampus tissue of the mice was significantly increased (P<0.05).Compared with ASF-AE group, the difference in sleep latency in ASF-AE+FLU group was not statistically significant(P>0.05),the sleep time was significantly shortened(P<0.01), and the GABA level in the hippocampus tissue of the mice was significantly decreased(P<0.05). Conclusion: ASF-WE and ASF-AE better sedative and hypnotic effects, and their mechanism may be related to its involvement in regulating the activity of 5-HT nervous system and GABA nervous system.
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Effect of vitamin D receptor activation on hepatic fibrosis induced by bile duct ligation in mice and its mechanism
YANG Fan, LI Lihua
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  722-727.  DOI: 10.13481/j.1671-587x.20200409
Abstract ( 4 )   PDF (1188KB) ( 5 )  
Objective: To investigate the protective effect of vitamin D receptor (VDR) activation on the bile duct ligation (BDL)-induced liver fibrosis in the mice, and to elucidate its possible mechanism. Methods: Sixty male C57BL/6 mice were randomly divided into sham operation group, sham operation+ paricalcitol group, model group (BDL) and treatment group (BDL+paricalcitol) (n=15).In sham operation, the abdominal cavities were opened but the bile duct was not ligated. In model group, the bile ducts of the mice were ligated with double lines and the middle of the bile duct was severed. The mice in sham operation+paricalcitol group and treatment group were intraperitoneally injected with the VDR agonist paricalcitol (200 ng·kg-1, three time a week) 3 d before the operation and continued treatment for 5 d. The livers of mice were removed on the 5th day after bile duct ligation,and the morphology of liver tissue and the fibrosis degree were observed by HE staining and Sirus red staining. The levels of reactive oxygen species (ROS) in the liver tissue of the mice were detected by dihydroethidine (DHE) staining. Western blotting method was used to detect the expression levels of silence mating type information regulation 3 (Sirt3), 8-hydroxy guanine DNA glycosidase 1(OGG1), alpha-smooth muscle (α-SMA)and collagen Ⅰ (Col Ⅰ) in the liver tissue of the mice. Results: The HE staining results showed that the structure of liver cells was complete, no inflammatory cell infiltration; in model group, a large number of infiltrated inflammatory cells and spot-like focal necrotic areas were observed in the liver of the mice; compared with model group, the area of liver tissue necrosis in treatment group was significantly reduced and the infiltration of inflammatory cells was significantly decreased. The Sirus red staining results showed that there was only a small amount of fibrosis around the big bile duct in the liver tissue of the mice in sham operation group and sham operation+paricalcitol group; collagen deposition was obvious in the portal area of the liver tissue of the mice in model group;the collagen deposition around the bile duct of the mice in treatment group was reduced compared with model group.The DHE staining results showed that there was only a small amount of red fluorescence in the portal area in the liver tissue of the mice in sham operation group and sham operation+paricalcitol group; in model group, there were significantly more red fluorescence in the portal area and the area around the bile duct; the intensity of red fluorescence in level treatment group was lower than that in model group.The Western blotting results showed that the expression level of Sirt3 protein in liver tissue of the mice in model group was significantly lower than sham operation group and sham operation+paricalcitol group (P<0.05);the expression levels of OGG1, α-SMA and Col Ⅰ proteins were increased significantly(P<0.05);compared with model group, the expression level of Sirt3 protein in liver tissue of the mice in treatment group was significantly increased(P<0.05),and the expression levels of OGG1, α-SMA and Col Ⅰ proteins were increased obviously (P<0.05). Conclusion: VDR activation can reduces oxidative stress-induced hepatic stellate cell activation by increasing the Sirt3 protein; thereby reduces the BDL-induced liver fibrosis in the mice.
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Analysis on dose-effect relationship of Atractylodes Rhizoma dryness effect based on regulation of SCF and c-kit mRNA expressions in colon tissue
XIE Ying, ZHOU Zhongshi, LIU Yanju, XIAO Kunyu, LEI Lin
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  728-732.  DOI: 10.13481/j.1671-587x.20200410
Abstract ( 3 )   PDF (1536KB) ( 5 )  
Objective: To analyze the dose-effect relationship of dryness effect of Atractylodes Rhizoma based on the expressions of stem cell factor (SCF) and its receptor c-kit mRNA in colon tissue and to ensure the basic dose of Atractylodes Rhizoma which caused the dryness reaction in the mice,and to provide the reference for clinical medicine. Methods: Seventy-five C57BL/6 mice were divided into control group,185, 370, 555, and 740 mg·kg-1 Atractylodes oil groups, 15 mice in each group, and the administration volume was 0.015 mL·g-1;the mice were administered intragastrically for 14 d,and the mice in control group were given the same volume of 1% Tween-20 solution. The average daily water intake and daily average urine output of the mice were detected. Immunohistochemical method was used to detect the expression levels of aquaporin-2 (AQP2) in the kidney tissue of the mice;the sleen and kidney coefficients of the mice were calculated, and RT-PCR method was used to determine the expression levels of SCF and c-kit mRNA in the colon tissue; the dryness effects of the mice in various groups were investigated according the obove indicatros. Results: Compared with control group, the average daily water intakes, daily urine outputs,the AQP2 expression levels in kidney tissue,the kidney coefficients and spleen coefficients of the mice in 185 and 370 mg·kg-1 Atractylodes oil groups had no significant differences (P>0.05); the expression levels of SCF mRNA in colon tissue of the mice were significantly increased (t=4.859, P<0.01; t=6.651, P<0.05); the expression level of c-kit mRNA in colon tissue of the mice in 185 mg·kg-1 Atractylodes oil group was significantly increased (t=2.946, P<0.05), but in 370 mg·kg-1 Atractylodes oil group had no statistically significant difference(P<0.05).Compared with control group, the average daily urine output and the expression level of c-kit mRNA in colon tissue of the mice in 555 mg·kg-1 Atractylodes oil group were significantly reduced (t=7.708, P<0.01; t=8.465, P<0.01), but there were no statistically significant differences in other indicators (P>0.05). Compared with control group, the water intake and the AQP2 protein expression level in kidney tissue of the mice in 740 mg·kg-1 Atractylodes oil group were significantly increased (t=3.554, P<0.01; t=4.636, P<0.01),and the average daily urine volume, the spleen coefficient,and the expression levels of SCF and c-kit mRNA in colon tissue of the mice were decreased significantly (t=7.708, P<0.01; t=4.985, P<0.01; t=7.314, P<0.01; t=28.45, P<0.01). Conclusion: Low doses (185 and 370 mg·kg-1) of Atractylodes oil show no obvious dryness effect in the healthy mice, and the high doses(555 mg·kg-1 and above) of Atractylodes oil may show obvious dryness effect by influencing the SCF/c-kit signal pathway.
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Effect of probiotics on expression of IL-17 in periodontal tissue of estrogen-deficient mice induced by ovariectomy and its significance
HUANG Jiangyong, LI Chanxiu, ZHENG Zhichao, WANG Hongjuan, GUO Lyuhua, WU Zhe, LUO Tao
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  733-738.  DOI: 10.13481/j.1671-587x.20200411
Abstract ( 5 )   PDF (1403KB) ( 4 )  
Objective: To explore the effect of probiotic feeding on the interleukin-17 (IL-17) expression in periodontal tissue of the estrogen-deficient mice, and to elucidate the possible mechanism of the correlation between estrogen deficiency and the periodontitis susceptibility. Methods: A total of 15 female C57BL/J6 mice(8-week-old) were randomly divided into sham operation group, ovariectomy group and probiotics group (probiotics feeding after ovariectomy) (n=5). The bone mineral densities(BMD) of the mice in various groups before and after experiment were measured by mean bone density measurement technique and the IL-17 mRNA expression levels in bone marrow cells and gingiva tissus of the mice were determined by RT-qPCR. The percentages of Th17 cells in CD4+TCRβ+ B cells in bone marrow of the mice were measured by flow cytometry. Results: Compared with sham operation group, the BMD of femur of the mice in OVX group was significantly decreased (P<0.01), the expression levels of IL-17 mRNA in bone marrow cells and gingiva tissue were increased (P<0.01), and the percentage of CD4+TCRβ+Th17 cells in the bone marrow was increased(P<0.05).Compared with OVX group, the BMD of femur of the mice in probiotics group was significantly increased (P<0.01), the expression levels of IL-17 mRNA in bone marrow cells and gingiva tissue were decreased (P<0.01), and the percentage of CD4+TCRβ+Th17 cells in the bone marrow was decreased(P<0.05).Compared with sham operation group, the BMD of femur, the expression levels of IL-17 mRNA in bone marrow cells and gingiva tissue, and the percentage of CD4+TCRβ+Th17 cells in the bone marrow in probiotics group had no significant differences (P>0.05). Conclusion: Probiotics feeding can inhibit the upregulation of IL-17 mRNA level of the mice induced by estrogen deficiency, and has the preventive effect on the estrogen deficiency-induced periodontitis.
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Effects of macrophage stimulating 1 receptor inhibitor BMS777607 on proliferation and apoptosis of lung cancer cells
JIA Daqi, KONG Wencong, SU Rongjian, DU Xiaoyuan, HE Wubin
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  739-744.  DOI: 10.13481/j.1671-587x.20200412
Abstract ( 7 )   PDF (1274KB) ( 7 )  
Objective: To investigate the effects of macrophage stimulating 1 receptor(MSTIR) inhibitor BMS777607 on the proliferation and apoptosis of lung cancer A549 cells, and to clarify their possible mechanisms. Methods: The A549 cells of human lung cancer were selected and treated with BMS777607 at different concentrations (1, 5, 10, 15,and 20 μmol·L-1) for 48 h;at the same time,control group was set up.The proliferation rates of A549 cells in various groups were detected by MTT method.The A549 cells were treated with BMS777607 at different concentrations (1, 5, 10, 15,and 20 μmol·L-1) for 14 d;at the same time,control group was set up.The colony formation of A549 cells in various groups were observed by colony formation experiment,and the proliferation rates of A549 cells were detected.The A549 cells were treated with different concentrations(10 and 20 μmol·L-1) of BMS777607 for 24 h;at the same time,control group was set up.The number of EdU positive cells in various groups were detected by EdU incorporation method. The A549 cells were treated with BMS777607 at different concentrations (1, 5, 10, 15, 20μmol·L-1) for 48 h;at the same time,control group was set up.The apoptotic rates were detected by flow cytometry.Western blotting method was used to detect the expression levels of protein kinase B(AKT),phosphorylated AKT (p-AKT), extracellular regulatory protein kinase(ERK),phosphorylated ERK(p-ERK), polyglycoside diphosphate ribose polymerase (PARP), Cleaved-PARP, Caspase 9 and Cleaved-Caspase 9. Results: The MTT assay results showed that compared with control group, the proliferation rates of A54 cells in different concentrations of BMS777607 groups were decreased significantly (P<0.05 or P<0.01) in a concentration and time dependent manner.In colony formation experiment,compared with control group,the number of colony formation of A549 cells in 10 μmol·L-1 BMS777607 group was significantly decreased,and the colony formation almost was not found in 20 μmol·L-1 BMS777607 group; compared with control group,the colony formation rates in different concentrations of BMS777607 groups were significantly decreased (P<0.05 or P<0.01).In experiment of EdU incorporation, the proliferation rates of A549 cells in 10 and 20 μmol·L-1 BMS777607 groups were decreased significantly compared with control group (P<0.05 or P<0.01).The flow cytometry results showed that compared with control group, the apoptotic rates of A549 cells in 10, 15 and 20 μmol·L-1BMS777607 groups were significantly increased (P<0.05).The results of Western blotting method showed that the expression levels of p-AKTand p-ERK proteins in the A549 cells in different concentrations of BMS777607 were significantly decreased(P<0.05), and the expression levels of Cleaved-PARP and Cleaved-Caspase 9 were significantly increased (P<0.05). Conclusion: MST1R inhibitor BMS777607 can inhibit the proliferation of lung cancer A549 cells and induce apoptosis,and its mechanism may be related to the inhibition of the expressions of p-AKT and p-ERK and the promotion of the expressions of PARP and Caspase 9.
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Construction of eukaryotic expression vector of human COX1 protein and its bioinformatics analysis
DONG Yuan, LI Jianhua, JIA Junzhen, FENG Wenzhuo, MENG Chenyang, HE Jiamei, TANG Yixin, WANG Huiyan
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  745-750.  DOI: 10.13481/j.1671-587x.20200413
Abstract ( 3 )   PDF (1534KB) ( 10 )  
Objective: To construct the eukaryotic expression vector of cycxygenase 1 (COX1) and to verify that COX1 protein can catalyze the synthesis of prostaglandin E1 (PGE1) in vitro, and to find a method to efficiently obtain PGE1. Methods: The total RNA of human microvascular endothelial cells (HMECs) was extracted and reversely transcribed into cDNA, which was served as the template to amplify the human COX1 gene by PCR. After double enzyme digestion, COX1 gene was connected with the eukaryotic expression vector pCMV6 to construct the recombinant plasmid pCMV6-COX1. Bioinformatic online softwares were used to analyze the hydrophobicity, transmembrane region, signal peptide, secondary structure and tertiary structure of COX1 protein. Afterwards, the human COX1 eukaryotic expression plasmid was expressed in the HEK-293F cells by liposomes. The cells were divided into control group (the HEK-293F cells without recombinant plasmid transfection), transfection 2 d group and transfection 5 d group. Then the cells and the cell culture supernatants were collected, respectively; the expression levels of COX1 protein were detected by Western blotting method. The activities of PGE1 synthesized by dohomo-γ-linolenic acid catalyzed by the eukaryotic expression COX1 protein and crude enzyme extract from sheep seminal vesicles were compared. Results: The recombinant plasmid pCMV6-COX1 was successfully constructed and identified by PCR, double digestion and sequencing. The length of target gene was about 1 800 bp. The bioinformatics analysis results showed that COX1 protein was a water-soluble protein. The amino acids at position 1-53 were signal peptides, the amino acids at position 34-53 were transmembrane regions; there were 36 serine phosphorylation sites, 14 threonine phosphorylation sites and 9 tyrosine phosphorylation sites. The prediction of secondary structure showed that the ratio of irregular crimp, alpha helix, extension chain and β angle were 46.20%, 41.03%, 10.18% and 2.58%, respectively.The Western blotting results showed that the recombinant plasmid was successfully transfected into the HEK-293F cells; compared with transfection 2 d group,the expression level of COX1 protein in the supernatants of the cells in transfetion 5 d group was significantly increased(P<0.05) and the relative molecular weight of protein was 70 000. When the substrate concentration was 1%, the yield of PGE1 synthesized by COX1 protein was (50.01±1.37) ng·L-1, which was equivalent to (90.3±0.06)% of the activity of crude enzyme extracted from 5 sheep seminal vesicles. Conclusion: The COX1 protein constructed and expressed by genetic engineering technology can catalyze the synthesis of PGE1 and meet the clinical requirements.
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Inhibitory effect of sileneing Frat2 on NCI-H1688 cells and its Wnt/β catenin signaling pathway mechanism
CHEN Yan, PAN Dianzhu, LIU Zhong, MA Yanmei, ZHANG Lan
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  751-758.  DOI: 10.13481/j.1671-587x.20200414
Abstract ( 6 )   PDF (1947KB) ( 6 )  
Objective: To investigate the effect of frequently rearranged in advanced T-cell lymphomas 2 (Frat2) on the proliferation of small cell lung cancer NCI-H1688 cells via Wnt/β-catenin signaling pathway,and to elucidate its possible mechanism. Methods: The human small cell lung cancer NCI-H1688 cells with good growth status and the normal bronchial epithelial-like cells HBE were selected;Western blotting method and Real-Time RT-qPCR were used to detect the expression levels of Frat2 mRNA and protein in the NCI-H1688 cells and the HBE cells.The NCI-H1688 cells were randomly divided into blank control group, negative control group,Frat2-siRNA group and Frat2-siRNA+XAV group.The cells in blank control group were not transfected.The cells in negative control group were transfected with negative control lentivirus,the cells in Frat2-siRNA group were transfected with Frat2-siRNA lentivirus and the cells in Frat2-siRNA+XAV group were transfected with the Frat2-siRNA lentivirus and treated with 4 μmol·L-1of Wnt signaling pathway inhibitor XAV939.The expression levels of Frat2 protein and mRNA in the cells in various groups were determined by Western blotting method and Real-Time RT-qPCR method.The proliferation activities of the cells in various groups were determined by MTT method.The percentages of cells in different cell cycles were measured by flow cytometry.The expression levels of Frat2, proliferating cell nuclear antigen (PCNA), c-myc, cyclin D1, β-catenin and phosphorylated glycogen synthase kinase 3 (pGSK-3β) proteins in the cells were determined by Western blotting method. Results: Compared with the normal bronchial epithelial-like cells HBE, the levels of Frat2 protein and mRNA in the NCI-H1688 cells were elevated (P<0.01).Compared with blank control group and negative control group, the expression levels of protein and Frat2 mRNA in Frat2-siRNA group were increased (P<0.05).There were no significant difference in the expression levels of Frat2 protein and mRNA in the NCI-H1688 cells between blank control group and negative control group (P>0.05).Compared with blank control group and negative control group, the proliferation activity of the cells in Frat2-siRNA group was decreased (P<0.05), the percentage of cells in G0/G1 phase was increased (P<0.05),the percentages of cells in S phase and G2/M phase were decreased (P<0.05), and the expression levels of PCNA, c-myc, Cyclin D1, β-catenin and pGSK-3β proteins were decreased (P<0.05).Compared with Frat2-siRNA group,the proliferation activity of cells in Frat2 mRNA+XAV group was decreased(P<0.05),the percentage of cells in G0/G1 phase was increased (P<0.05), the percentages of cells in S phase and G2/M phase were decreased (P<0.05),and the expression levels of PCNA, c-myc, Cyclin D1, β-catenin and pGSK-3β proteins were decreased (P<0.05).There were no significant differences in the indexes mentioned above between blank control group and negative control group (P>0.05). Conclusion: Silencing Frat2 can inhibit the proliferation of small cell lung cancer cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.
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Expression of insulin-like growth factor binding protein-3 in ischemic brain tissue of cerebral infarction model rats and its relationship with angiogenesis
BU Yi, ZHANG Shuo, QIAN Xudong, WANG Hongmei, DOU Zhijie
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  759-764.  DOI: 10.13481/j.1671-587x.20200415
Abstract ( 2 )   PDF (1527KB) ( 3 )  
Objective: To investigate the expression level of insulin-like growth factor binding protein-3(IGFBP-3) in ischemic brain tissue of the rats with cerebral infarction and its relationship with angiogenesis,and to elucidate the role of IGFBP-3 in angiogenesis of cerebral infarction. Methods: The SD rats were randomly divided into control group and model group,and the rats in model group were divided into three subgroups of model 1 d group, model 3 d group and model 7 d group, with 30 rats in each group.The rat model of middle cerebral artery occlusion ischemia was established by suture method.Neurobehavioral score method was used to assess the neural behavior of the rats.Cerebral infarctions were observed by 2,3,5-triphenyltetrazolium chloride (TTC) staining.The pathomorphology of brain tissue of the rats was observed by HE staining.Immunohistochemical staining was used to measure the expression levels of IGFBP-3 and vascular endothelial growth factor (VEGF) proteins in brain tissue of the rats.Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression levels of IGFBP-3 and VEGF mRNA in brain tissue of the rats.Immunofluorescence staining was used to measure the microvessel density in the ischemic penumbra of the brain tissue of the rats. Results: There were no neurological deficits and cerebral infarction of the rats in control group.In model group,the neurobehavioral score of the rats was (1.86±0.73) scores and the volume of cerebral infarction was (28.34±3.57)%.The HE staining results showed that the brain tissue of the rats in control group was normal;in model 1 d group,edema occurred in the brain tissue of the rats; in model 3 d group,edema and necrosis of brain tissue of the rats were obvious; in model 7 d group, edema and necrosis of brain tissue were alleviated.Compared with control group, the expression levels of IGFBP-3 and VEGF proteins and mRNA in brain tissue of the rats in various model groups and the microvessel densities in ischemic penumbra were increased (P<0.01);the above indexes in brain tissue of the rats in model 3 d group were the highest;the expression levels of IGFBP-3 and VEGF proteins and mRNA in brain tissue of the rats in model 7 d group and the microvessel density in ischemic penumbra were decreased.There were positive correlation between the expression levels of IGFBP-3 mRNA and the VEGF mRNA and microvessel density in brain tissue of the rats (r=0.563,P<0.05;r=0.612,P<0.05). Conclusion: The expression level of IGFBP-3 in ischemic brain tissue of the rats with cerebral infarction is increased, which may promote angiogenesis after cerebral infarction.
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Protective effect of paeonol on myocardial injury in rats with heart failure and its mechanism
GAO Ruimin, KANG Lingling, HOU Fangzhen
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  765-770.  DOI: 10.13481/j.1671-587x.20200416
Abstract ( 8 )   PDF (2265KB) ( 7 )  
Objective: To investigate the protective effect of paeonol on myocardial injury in the rats with heart failure, and to elucidate its preliminary pharmacological mechanism. Methods: The rat models of heart failure were replicated by intraperitoneal injection of doxorubicin. After successful modeling, the rats were randomly divided into model group(n=10), captopril group(n=11), low dose of paeonol group(n=11) and high dose of paeonol group(n=11), and the normal rats were used as control group(n=10).After successful modelring, the rats were given the drug continuously for 5 weeks, and the body weights of rats were recorded.After the last administration,the rats were anesthetized,and the changes of mean arterial pressure(MAP),heart rate(HR),left ventricular end diastolic pressure(LVEDP), left ventricular systolic blood pressure (LVSP), maximum rate of decrease of left ventricular blood pressure (-dp/dtmax), and maximum rate of increase of left ventricular blood pressure (+dp/dtmax)]of the rats were observed.The serum of rat orbital peripheral blood was isolated, and the levels of the serum tumor necrosis factor (TNF-β) and interleukin-6 (IL-6) were detected by ELISA. After anesthesia, 5 rats were randomly selected and the heart was obtained under aseptic operation; Western blotting method was used to detect the expression levels of Bax, B cell lymphoma - 2 (Bcl-2), cysteine aspartic acid protease 3 (Caspase-3), cysteine aspartic acid protease 8 (Caspase-8), cysteine aspartic acid protease 9 (Caspase-9) and cytochrome C (Cyt C) proteins in myocardium tissue of the rats in various groups. HE staining was used to observe the pathomorphology of myocardium tissue of the rats. Results: Compared with control group, the body weight of the rats in model group was significantly reduced (P<0.05),and MAP and LVEDP were increased(P<0.05), HR, LVSP, +dp/dtmax and -dp/dtmax were significantly decreased (P<0.05),the serum TNF-α and IL-6 levels were increased (P<0.01), the expression levels of Bax,Caspase-3,Caspase-8,Caspase-9 and Cyto-C proteins were significantly increased (P<0.01),and the expression level of Bcl-2 was significantly decreased (P<0.05). Compared with model group, the body weights of the rats in captopril,low and high doses of paeonol groups were significantly increased (P<0.05),the cardiac function indexes were improved significantly (P<0.05), the TNF-α and IL-6 levels were decreased (P<0.05), the expression levels of Bax, Caspase-3, Caspase-8, Caspase-9 and Cyto-C proteins in myocardium tissue of the rats were significantly decreased (P<0.05), and the Bcl-2 protein expression levels were increased significantly (P<0.05).The HE staining results showed that compared with control group, the myocardium tissue of the rats in model group was damaged obviously, and the myocardial cells were edema extensively. Compared with model group, the myocardial fiber arrangement and myocardial interstitial edema of the rats in captopril group and low and high doses of paeonol groups were significantly improved. Conclusion: Paeonol has a protective effect on the myocardium of the rats with heart failure, and the mechanism may be related to the improvement of hemodynamics, anti-inflammation and anti-apoptosis, and the protection of myocardial cells. High dose of paeonol is more effective in protecting the cardiac function and anti-apoptosis.
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Improvement effect of eleutheroside B on learning and memory abilities of fatigue mice and its mechanism of activating Keap1/Nrf2/ARE signaling pathway
ZHANG Qian, FENG Qingxia, ZHOU Zhengyi, LI Ruiqi, FANG Lei, YANG Qiunan, SHENG Yu, LU Xuechun, DU Peige, AN Liping
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  771-778.  DOI: 10.13481/j.1671-587x.20200417
Abstract ( 7 )   PDF (1540KB) ( 10 )  
Objective: To investigate the effect of eleutheroside B (ELB) on the learning and memory abilities in the fatigue mice, and to clarify its possible mechanism. Methods: A total of 60 ICR mice were randomly divided into control group (received distilled water by gavage, meditation experiment), model group (received distilled water by gavage, swimming experiment), ELB (C) group (received 80 mg·kg-1 ELB by gavage, meditation experiment) and ELB (M) group (received 80 mg·kg-1 ELB by gavage, swimming experiment) (n=15). After 4 weeks of continuous administration, the anti-fatigue abilities of the mice were observed with rotating rod experiment, and the learning and memory abilities of the fatigue mice were detected by morris water maze experiment and dark avoidance experiment; the levels of lactic acid (LD) in serum of the mice were detected by colorimetry, the activities of lactate deoxygenase (LDH) in serum of the mice were detected by microenzyme labeling method, the levels of urea nitrogen (BUN) in serum of the mice were detected by urease method, the activities of superoxide dismutase (SOD) in brain tissue of the mice were detected by hydroxylamine method,and the levels of malondialdehyde (MDA) in brain tissue of the mice were detected by thiobarbituric acid method; the levels of reactive oxygen species (ROS), endothelial nitric oxide synthase (eNOS), γ-aminobutyric acid (GABA) and glutamate (GLU) in brain tissue of the mice were detected by enzyme linked immunosorbent assay; the activities of adenosine triphosphase (Na+K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, Ca2+Mg2+-ATPase) in brain tissue of the mice were detected by spectrophotometry.Western blotting method was used to detect the expression leves of Kelch-like ECH-associated protein-1 (Keap1),nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins in brain tissues of the mice. HE staining was used to observe the structures of hippocampal neurons in brain tissue of the mice. Results: Compared with control group, the rotating rod time of the mice in model group was shortened (P<0.05), the water maze latency was extended (P<0.05), the dark avoidance latency was shortened (P<0.05), and the number of errors was increased (P<0.05); the activity of serum LDH and the levels of serum LD and BUN were increased (P<0.05); the level of eNOS, the activities of SOD, Na+K+-ATPase, Mg2+-ATPase and Ca2+-ATPase, Ca2+Mg2+-ATPase, and the GLU/GABA ratio in brain tissue of the mice were decreased (P<0.05), and the MDA and ROS levels were increased (P<0.05); the Keap1 protein expression level was increased (P<0.05),and the Nrf2 and HO-1 protein expression levels were decreased (P<0.05); the rotating rod time of the mice in ELB (C) group was prolonged(P<0.05), but the differences in the other indicators had no statistical significance (P>0.05).Compared with model group, the rotating rod time of the mice in ELB (M) group was prolonged(P<0.05), the water maze latency was shortened (P<0.05), the dark avoidance latency was prolonged (P<0.05), and the number of errors was reduced (P<0.05); the activity of serum LDH and the levels of serum LD and BUN were decreased (P<0.05), the level of eNOS and the activities of SOD, Na+K+-ATPase, Mg2+-ATPase and Ca2+-ATPase, Ca2+Mg2+-ATPase,and the GLU/GABA ratio in brain tissue of the mice were increased (P<0.05), and the MDA and ROS levels were decreased (P<0.05);the Keap1 protein expression level was decreased (P<0.05),and the Nrf2 and HO-1 protein expression levels were increased (P<0.05).Compared with control group, the arrangement of neurons in the hippocampus of the mice in model group was loose and unclear, the morphology of some hippocampal cells was changed, and there were obvious pathological changes.Compared with model group, the structures of the hippocampus region of the mice in ELB(C) and ELB(M) groups were clear, the number of neurons was increased,the arrangement was relatively tight and orderly, and the morphology and volume of hippocampal neurons tended to be normal. Conclusion: ELB can improve the learning and memory abilities of the fatigue mice, and its mechanism may be related to regulating the expressions of Keap1/Nrf2/ARE signaling pathway-related factors.
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Effect of berberine hydrochloride on exosomes of cervical cancer HeLa cells and its mechanism
WANG Jing, GE Jing, WANG Xu, FANG Guiying, DAI Lili, LIU Jing
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  779-785.  DOI: 10.13481/j.1671-587x.20200418
Abstract ( 14 )   PDF (1375KB) ( 5 )  
Objective: To observe the intervention effect of berberine hydrochloride on the exosomes released by cervical cancer HeLa cells, and to explore its effects on the abilities of proliferation and metastasis of cancer cells. Methods: The cervical cancer HeLa cells were divided into control group and low, medium and high doses of berberine hydrochloride groups, and they were intervented with 0, 10, 20 and 40 mg·L-1berberine hydrochloride for 24 h. High-speed centrifugation was used to extract the exosomes and Western blotting method was used to detect the expression levels of exosome markers CD81, CD63 and phosphatidylinositol 3-kinase(PI3K)/protein-serine-threonine kinase(Akt) signaling pathway related proteins. MTT assay was used to determine the inhibitory rates of growth of the HeLa cells.Scratch assay was used to measure the migration rate of cells. Transwell assay was used to detect the number of transmembrane cells. Results: Compared with control group, the expression levels of cervical cancer cell exosome markers CD81 and CD63 proteins in the exosomes in low, medium and high doses of berberine hydrochloride groups were significantly decreased (P<0.05), while the inhibitory rates of growth of cancer cells were increased significantly (P<0.05); the migration rates of cells and the number of transmembrane cells were decreased significantly (P<0.05),and the expression levels of PI3K, Akt and phosphorylated Akt (p-Akt) proteins in the cancer cells were significantly decreased (P<0.05).Compared with low dose of berberine hydrochloride group, the expression levels of CD81 and CD63 proteins in the exosomes in medium and high doses of berberine hydrochloride groups were significantly decreased (P<0.05), while the inhibitory rates of growth of cancer cells were increased significantly (P<0.05); the migration rates of cells and the number of transmembrane cells were decreased significantly (P<0.05), and the expression levels of PI3K, Akt and p-Akt proteins in the cancer cells were significantly decreased (P<0.05).Compared with medium dose of berberine hydrochloride group, the expression levels of CD81 and CD63 proteins in the exosomes in high dose of berberine hydrochloride group were significantly decreased (P<0.05), while the inhibitory rate of growth of cancer cells were increased significantly (P<0.05); the migration rates of cells and the number of transmembrane cells were decreased significantly (P<0.05), and the expression levels of PI3K, Akt and p-Akt proteins in the cancer cells were significantly decreased (P<0.05) in a concentration-dependent manner. Conclusion: Berberine hydrochloride can inhibit the exosome secretion of cervical cancer cells and the proliferation of cancer cells, and reduce the the abilities of invasion and metastasis of cancer cells. Its mechanism may be related to the down-regulation of the expressions of the PI3K/Akt signaling pathway related proteins.
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Effects of EZH2 inhibitor GSK126 on proliferation and apoptosis of lung adenocarcinoma cells
ZHANG Shuangshuang, HE Wubin, SU Rongjian, DU Xiaoyuan
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  786-791.  DOI: 10.13481/j.1671-587x.20200419
Abstract ( 1 )   PDF (1293KB) ( 6 )  
Objective: To investigate the effects of GSK2816126(GSK126), a specific inhibitor of zeste gene enhancer homolog 2 (EZH2), on the proliferation and apoptosis of lung adenocarcinoma A549 cells, and to elucidate their possible mechanisms. Methods: The A549 cells were cultured in vitro and divided into control group and different concentrations(1.0,2.5,5.0,10.0,15.0 μmol·L-1)of GSK126 groups. MTT colorimetry was used to detect the survival rates of cells in various groups.Clone formation experiment was used to detect the number of colony formation and the colony formation rate was calculated. EdU imaging was used to detect the proliferation rates of cells in various groups. Hoechst-33342 fluorescence staining was used to observe the apoptotic morphology of cells in various groups; flow cytometry was used to detect the apoptotic rates of A549 cells in various groups. Western blotting method was used to detect the expression levels of protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), B-cell lymphoma-2 proto oncogene (Bcl-2), Bcl-2 related X protein (Bax), aspartate specific cysteine proteinase-3 (caspase-3) and activated aspartate specific cysteine proteinase-3 (Cleaved-Caspase-3) proteins in the cells in various groups. Results: The MTT results showed that compared with control group, the survival rates of A549 cells in different concentrations of GSK126 groups were decreased with the increase of GSK126 concentration (P<0.05).The results of clone formation experiment showed that compared with control group, the number of colonies in different concentrations of GSK126 groups were decreased gradually with the increase of GSK126 concentration (P<0.05).The EdU imaging results showed that the proliferation rates of the cells in different concentrations of GSK126 groups were decreased significantly with the increase of GSK126 concentration compared with control group (P<0.01).The Hoechst-33342 staining and flow cytometry results showed that compared with control group, the apoptotic rates of the cells in different concentrations of GSK126 groups were increased significantly with the increase of GSK126 concentration (P<0.01).The Western blotting results showed that compared with control group, the expression levels of p-AKT and Bcl-2 proteins were decreased gradually with the increase of GSK126 concentration (P<0.05), and the expression levels of Bax and Cleaved-Caspase-3 proteins were increased gradually (P<0.05). Conclusion: GSK126 may inhibit the cell proliferation by reducing the phosphorylation level of AKT, and promote the apoptosis of lung cancer cells by inducing Bax/Bcl-2/Caspase-3 signal pathway activation and activating apoptosis pathway.
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Protective effect of sufentanil post-conditioningthrough ERK1/2-mediated p70S6K signaling pathway on myocardial ischemia-reperfusion injuryof rats
WEN Ying, WU Qiaoling, LIU Guoli
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  792-797.  DOI: 10.13481/j.1671-587x.20200420
Abstract ( 5 )   PDF (1011KB) ( 5 )  
Objective: To investigate the effect of sufentanil post-conditioning on the myocardial ischemia reperfusion injury (MIRI) through extracellular regulated protein kinases (ERK1/2)-mediated p70S6K signaling pathway in the rats, and to clarify the protective effect of sufentanil post-conditioning on the MIRI of the rats. Methods: Seventy-two male SD rats were randomly divided into sham operation group, ischemia-reperfusion group (I/R group), dimethylsulfoxide group (DMSO group), PD group (an inhibitor of ERK, 20 μmol·L-1 PD98059 was administered 15 min after the end of ischemia to the start of reperfusion), sufentanil post-conditioning group (Sufen group), Sufen+PD group. Except for sham operation group, the rats in other groups were reperfused for 2 h after 30 min-ischemia. The heart rates (HR) and mean artery pressures (MAP) of the rats in various goups were recorded immediately before ischemia (T0), 30 min of ischemia (T1), 30 min of reperfusion (T2), 60 min of reperfusion (T3), 90 min of reperfusion (T4), and 120 min (T5). The myocardial infarction area was measured at the end of reperfusion; the expression levels of p-ERK1/2 and p-p70S6K proteins in myocardium tissue were detected by Western blotting method. Results: Compared with T0, The MAP and HR of the rats in I/R, Sufen, DMSO, PD, and PD + Sufen groups at T1-T4 were decreased (P<0.05).Compared with sham operation group, the HR and MAP of the rats in other groups were increased(P<0.05). Compared with I/R group, the HR and MAP of the rats in Sufen group were increased (P<0.05).Compared with I/R group (45.67% ±10.32%), the myocardial infarct area of the rats in Sufen group (20.92% ±6.88%) was reduced (P<0.05).Compared with Sufen group, the myocardial infarction area in PD + Sufen group (42.14% ±6.73%) was increased (P<0.05). Compared with sham operation group, the expression levels of p-ERK1/2and p-p70S6K proteints in myocardium tissue of the rats in I/R group, Sufen group, and DMSO group were increased (P<0.05).Compared with I/R group, the expression levels of p-ERK1/2 and p-p70S6K in myocardium tissue of the rats in Sufen group were further increased(P<0.05).Compared with Sufen group, the expression levels of p-ERK1/2 and p-p70S6K proteins in myocardium tissue of the rats in PD+Sufen group were decreased(P<0.05). Conclusion: Sufentanil post-conditioning has the protective effect on MIRI in the rats through ERK1/2-mediated p70S6K signaling pathway. Sufentanil post-conditioning improves the cardiac function indicators and reduces the myocardial infarction area.
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Effect of HeLa cell exosomes on migration and invasion and its mechanism of Wnt/ β-catenin signaling pathway
LIU Jing, WANG Jing, GE Jing, FENG Yanping, FANG Guiying, WANG Xu, YANG Yanhong, LI Lin
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  798-803.  DOI: 10.13481/j.1671-587x.20200421
Abstract ( 7 )   PDF (2367KB) ( 6 )  
Objective: To investigate the effects of cervical cancer HeLa cell exosomes on the migration, invasion and Wnt/β-catenin signaling pathway of the cells, and to elucidate the possible mechanism of the exosomes in the occurrence and development of cervical cancer. Methods: The cervical cancer HeLa cells in the logarithmic growth phase were selected. The cervical cancer HeLa cell exosomes were isolated by ultracentrifugation. The morphology of isolated exosomes was observed under transmission electron microscope(TEM). Western blotting method was used to detect the expressions of exosomal marker proteins CD63 and CD81, and the isolated exosomes were identified. The HeLa cells were divided into exosomes group(Exo group) and control group. The cells in two groups were cultured separately with medium containg HeLa cell exosomes or not. The uptake of exosomes by HeLa cells was observed with confocal microscope, the exosomes were labeled with PKH26 fluorescence, and the nuclei of HeLa cells were stained with DAPI fluorescence. Transwell chamber was used to measure the cell invasion ability.Cell scratch assay was used to measure the cell migration ability.Western blotting method was used to measure the expression levels of Wnt1, β-catenin and T cell factor 4 (TCF4) proteins in the cells. Results: The TEM results showed cylindrical or oval vesicle bodies with a diameter of 40-100 nm and rich in CD63 protein and CD81 protein. The confocal microscope results showed that there was exosomal attachment on the membrane surface of HeLa cells in Exo group,but there was no exosomal attachment on the membrane surface of the HeLa cells in control group; The Transwell assay results showed that the number of invasion HeLa cells in Exo group was significantly higher than that in control group(t=17.894,P<0.01);the scratch assay results showed that the migration distance of the cells in Exo group was significantly longer than that in control group(t=9.689,P<0.01).The Western blotting results showed the expression levels of Wnt/β-catenin signaling pathway-related proteins Wnt1, β-catenin and TCF4 in the HeLa cells in Exo group were significantly higher than those in control group(t=13.159,P<0.01;t=15.478,P<0.01;t=7.734,P<0.01). Conclusion: The HeLa cell-derived exosomes can promote the invasion and migration abilities of cervical cancer cells. The mechanism may be related to that the HeLa cell-derived exosomes can promote the activation of Wnt/β-catenin signaling pathway and the downstream TCF4 gene expression.
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Research in clinical medicine
Analysis on endometrial cancer-related genes and candidate pathways based on GEO database bioinformatics methods
WANG Zhi, HONG Li, LI Suting, ZENG Wanling
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  804-809.  DOI: 10.13481/j.1671-587x.20200422
Abstract ( 4 )   PDF (1836KB) ( 7 )  
Objective: To analyze the key genes and candidate pathways related to the occurrence and development of endometrial cancer(EC) with the bioinformatics methods, and to explore the pathogenesis and the therapeutic targets of EC. Methods: The EC datasets (GSE17025 and GSE63678) were downloaded from the Gene Expression Omnibus (GEO), and the GEO2R online analysis tools and R software were used to screen for the differential expression genes (DEGs) in the EC tissue and the adjacent normal tissue. The GO enrichment analysis and KEGG pathway analysis of DEGs were performed with the String database for protein-protein interaction network (PPI) analysis. Finally, the PPI network was analyzed and visualized by Cytoscape software. Results: After the DEGs analysis of the datasets GSE17025 and GSE63678, 100 co-upregulated genes and 106 co-downregulated genes were obtained. The results of GO enrichment analysis indicated that DEGs were mainly enriched in mitotic chromosome segregation, nuclear division, organelle division and other biological processes. The result of KEGG signaling pathway analysis showed that DEGs were mainly enriched in cell cycle, miRNA, p53 signaling pathway, type Ⅱ diabetes signal pathway. Through Cytoscape software analysis,CDC20, AURKA, CCNB1, DTL, CEP55, CDK1, KIF11, MELK, CCNB2, and BUB1 in the PPI network were screened as the key genes. Conclusion: The imbalance of cell cycle-related genes and pathway regulatory networks may be involved in the occurrence of EC.
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Effect of preparation plan of hormone replacement after down-regulation on pregnancy outcome of frozen-thawed embryo transfer cycle in advanced age infertile patients
WANG Lei, YAN Xiaohong, LIN Li, LI Youzhu
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  810-815.  DOI: 10.13481/j.1671-587x.20200423
Abstract ( 5 )   PDF (1849KB) ( 8 )  
Objective: To investigate the effects of two methods of endometrial preparation plans(hormone replacement after down regulationand hormone replacement) on the pregnancy outcome of frozen-thawed embryo transfer(FET) cycle in the advanced age infertile patients, and to analyze the influencing factors of pregnancy outcome of FET cycle of hormone replacement after down-regulation and to provide the basis for the selection of endometrial preparation scheme for FET cycle in the aged infertile patients. Methods: A total of 211 patients aged ≥ 35 years who first received FET treatment were selected.The patients were divided into hormone replacement cycle after down-regulation group(n=114) and hormone replacement cycle group(n=97).The age, duration of infertility, body mass index (BMI), anti-Mullerian hormone(AMH) level, basic sex hormones, endometrial thickness on the day of conversion, number of embryos transferred, number of high-quality embryos transferred, rates of high-quality embryos transferred,rates of multiple pregnancies, and abortion rates of the patients in two groups were analyzed, retrospectively, and the influencing factors of pregnancy outcome of hormone replacement cycle after down-regulating were analyzed by Binomial Logistic regression. Results: There were no significant differences in age, duration of infertility,BMI, AMH levels, number of embryos transferred, number of high-quality embryos transferred, rates of high-quality embryos transferred, rates of multiple pregnancies, and cycle abortion rates of the patients between two groups(P>0.05).The clinical pregnancy rate and embryo implantation rate of the patients in hormone replacement cycle after down-regulating group were significantly higher than those in hormene replacement cycle group(χ2=9.964,P<0.05;χ2=9.964,P<0.05). The results of Logistic regression analysis showed that age (OR=0.805,95%CI:0.697-0.930) and the number of high-quality embryos transferred (OR=4.098,95%CI:1.597-10.514) were the key factors influencing the pregnancy outcome of hormone replacement cycle after down-regulation in the advanced age infertile patients. Conclusion: Hormone replacement cycle after down-regulation can achieve the better clinical outcomes in FET in the advanced age infertility patients. Age and number of high-quality embryos transferred are the key factors affecting the pregnancy outcome of hormone replacement cycle after down-regulation in the advance age infertile patients.
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Application of hematology-related indexes in early diagnosis of bacterial infectionof tumor patients in ICU
LI Ting, CHEN Yuanyuan, SUN Hongshuai, YU Xiuyan, WU Xuefeng
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  816-821.  DOI: 10.13481/j.1671-587x.20200424
Abstract ( 6 )   PDF (1846KB) ( 5 )  
Objective: To detect the levels of the hematology-related indexes in the tumor patients in intensive care unit(ICU), and to investgate their clinical application in the early diagnosis of bacterial infection in the tumor patients in ICU. Methods: The complete clinical materials of 256 tumor patients in ICU were analyzed retrospectively. According to the clinical bacterial infection standard, they were divided into infection group(67 cases) and non-infection group(189 cases), and according to the situation of death of patients, they were divided into survival group(193 cases) and death group(63 cases). The prothrombin time (PT), active part thrombin time (APTT) and D-Dimer(DD),the levels of procalcitonin (PCT) and the counts of white blood cell(WBC), neutrophile(NEU), lymphocyte(LYM) of the tumor patients in ICU in infection group and non-infection group were measured, and the neutrophile-to-lymphocyte ratio(NLR) was calculated. The sensitivities and specificities of each index in the early diagnosis of bacterial infection in the ICU tumor patients and PCT for predicting the mortality risk in the ICU tumor patients with early bacterial infection were assessed by receiver operating characteristic curve(ROC) and the area under the curve(AUC), and the their clinical values were explored. Results: Compared with non-infection group, the PT, APTT, DD, PCT level and NRL of the ICU tumor patients in infection group were significantly increased(P<0.05 or P<0.01).Compared with PCT<0.25 μg·L-1 group, the DD and NRL levels of the ICU tumor patients with early bacterial infection in 0.25 μg·L-1 ≤ PCT<2.00 μg·L-1 group,2.00 μg·L-1 ≤ PCT<10.00 μg·L-1 group and PCT ≥ 10.00 μg·L-1 group were significantly increased(P<0.05 or P<0.01); the PT and APTT levels of the ICU tumor patients with early bacterial infection in 2.00 μg·L-1 ≤ PCT<10.00 μg·L-1 group and PCT ≥ 10.00 μg·L-1 group were significantly increased(P<0.05).The AUC, sensitivities and specificities of PT, APTT, DD, NLR and PCT in the diagnosis of early bacterial infection in the ICU tumor patients were 0.636, 60.8%, 59.1%; 0.622, 64.6%, 58.1%;0.672, 69.6%, 58.1%;0.752, 74.7%, 65.6%; 0.855, 70.9%, 83.9%; the differences were statistically significant (P<0.01).The PCT level of ICU tumor patients with early bacterial infection in death group was significantly higher than that in survival group (P<0.05).The AUC of PCT for predicting the mortality risk of ICU tumor patients with early bacterial infection was 0.803(95%CI:0.749-0.857), the cut-off value was 6.72 μg·L-1, the sensitivity and specificity were 63.2% and 79.1%, respectively, and the differences were statistically significant (P<0.01). Conclusion: PT, APTT, DD, PCT and NLR can be used as the early diagnostic indicators of bacterial infection in the ICU tumor patients. The sensitivity of NLR and the specificity of PCT were the highest. PCT can be used as a predictor of the mortality risk in the ICU tumor patients with early bacterial infection.
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Effects of different root canal filing materals on root flexure resistance after random load fatigue test detected by Micro-CT
ZHANG Li, QUAN Tao, LIU Dayong, JIA Zhi
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  822-827.  DOI: 10.13481/j.1671-587x.20200425
Abstract ( 5 )   PDF (2609KB) ( 8 )  
Objective: To detect the root crack after in vitro fatigue test with Micro-computed tomography(Micro-CT), and to explore the effects of two different root canal filling materials on the root flexure resistance. Methods: A total of 60 freshly extracted human single canal anterior teeth were randomly divided into control group (n=20),OrthoMTA group (n=20), and Gutta-Percha + AH-plus group (n=20). After removing the crowns of all samples, the root canals were prepared using the BLX nickel titanium system. The root canals in control group were not filled,the root canals in OrthoMTA group were filled with OrthoMTA root canal,and the root canals Gutta-Percha + AH-plus group were filled with Gutta-Percha and AH-plus sealant. All samples were placed in a mechanical cycler (500-1500 N, 2 Hz, with sinusoidal alternation and cycle 7 200 times),and the in vitro fatigue test was performed.Micro-CT was used to detect the internal cracks in the roots of each group and the crack severity scores were evaluated. Results: The results of Micro-CT scanning showed that after stress, most of the roots in control group, OrthoMTA group and Gutta-Percha+AH-plus group appeared the cracks of varying degrees, including incomplete cracks, complete cracks, and two types of cracks. Most of the cracks were in the near, far, and middle directions.The crack score of the root in OrthoMTA group was lower than that in control group, but the difference was not statistically significant (P>0.05). The crack score of the root in Gutta-Percha + AH-plus group was higher than that in control group, but the difference was not statistically significant (P>0.05); the crack score of the root in OrthoMTA group was lower than that in Gutta-Percha + AH-plus group, and the difference was not statistically significant (P>0.05). Conclusion: The random load fatigue test confirms that the in vitro resistance model is constructed successfully. Based on this study, OrthoMTA complete root canal filling and Gutta-Percha + AH-plus filling have no significant difference in influence in the root flexure resistance.
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Comparison of biomechanical effects of repairing lower molar residual crowns among post-core crown, endocrown and inlaycrown and their clinical significances
LI Weixuan, JIN Julou, ZHAO Chuqiao, LIU Dingkun, ZOU Jundong, WANG Mingxia, LIU Zhihui
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  828-833.  DOI: 10.13481/j.1671-587x.20200426
Abstract ( 7 )   PDF (2256KB) ( 7 )  
Objective: To compare the stress distribution and retention effects of lower molar residual crowns restored by post-core crown, endocrown and inlaycrown using the finite element analysis, and to explore the optimized repair plan of lower molar residual crowns. Methods: The healthy and integral left mandibular first molar of volunteer was selected as the experimental sample. Their imaging data was collected by cone beam CT (CBCT) scanning. Then the complete finite element model of the mandibular first molar was reconstructed by reverse engineering software such as Mimics, Geomagic and CATIA. On this base, three groups of models of lower molar residual crowns with the clinical crown heights of 1, 2, and 3mm were constructed. Meanwhile, the finite element models of post-core crown, endocrown, and inlaycrown were established to repair the three groups of residual crowns mentioned above; there were nine experimental groups. In the Abaqus software, the vertical and oblique static loads were applied on the models to simulate the forces produced during chewing, and the forced rotational displacement load was applied on the models to simulate the dislocation of restorations. Then the stress peak and stress distribution cloud map of von Mises in dentin of the models in various groups, as well as the force generated by restorations to resist dislocation and the damage of adhesive during dislocating were all observed. Results: The von Mises stress peak results were inlaycrown > endocrown > post-core crown in vertical load and endocrown > inlaycrown > post-core crown in oblique load. According to the stress distribution cloud map, the root apical 1/3 of post-core crown, the pulp floor of endocrown and the dental cervix and root of inlaycrown was found obviously in the stress concentration. The non-axial retention force results were post-core crown > inlaycrown > endocrown in the 1 mm residual crown group, and inlaycrown > post-core crown > endocrown in the 2 and 3 mm residual crown groups. According to stiffness degradation contours of resin adhesive layer, the cracked areas of adhesive during dislocation was inlaycrown > endocrown > post-core crown. Conclusion: From the perspectives of protecting dental tissue and maintaining the stability of restorations, the post-core crown is an ideal restoration for repairing the lower molar residual crowns.
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Expressions of plasmacytoid dendritic cells and Foxp3+ regulatory T cells in colorectal cancer tissue and their significances
WANG Dan, LIAO Dan, LI Hong, XIONG Liqiu, WU Ying, DONG Ying, GAI Xiaodong
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  834-838.  DOI: 10.13481/j.1671-587x.20200427
Abstract ( 5 )   PDF (276KB) ( 7 )  
Objective: To investigate the expressions of CD123+ plasmacytoid dendritic cells(pDCs) and Foxp3+ regulatory cells(Tregs) in the colorectal cancer tissue (CRC) and their infiltrations in the tumor draining lymph nodes (TDLN), and to elucidate their effects on the biological behaviors of CRC. Methods: A total of 63 pathological specimens of removed CRC tissue, cancer adjacent normal mucosa tissue and TDLN were collected. The number of CD123+pDCs and Foxp3+Treg positive cells in different tissues were detected by immunohistochemical method. The correlation between the expressions of them were detected by Spearman analysis. Results: The number of Foxp3+Tregs positive cells in cancer tissue of the CRC patients was higher than that in adjacent normal mucosa tissue(P<0.01). The number of Foxp3+Tregs positive cells in cancer tissue of the patients with positive lymph node metastasis was higher than that in adjacent normal mucosa (P<0.01).The number of Foxp3+Tregs positive cells in metastatic TDLN (mTDLN) was higher than that in metastatic free TDLN (mfTDLN) (P<0.01).The number of Foxp3+Tregs psitive cells in cancer tissue in the patients in TNM Ⅲ+Ⅳstage was higher than that in the patients in TNM Ⅰ+Ⅱ stage (P<0.01).The number of CD123+pDCs positive cells in cancer tissue of the CRC patients was lower, and the positive expression rate of CD123 in CRC tissue was signifificantly higher than that in adjacent normal mucosa tissue (P<0.01).The number of CD123+pDCs positive cells in mfTDLN and mTDLN had no significant difference (P>0.05).The pDCs/myeloid dendritic cells(mDCs) ratio in mTDLN was signifificantly higher than that in mfTDLN(P<0.01).The correlative analysis results demonstrated that the ratio of pDCs/mDCs had a positive relationship with the number of Foxp3+ Tregs positive cells(r=0.421, P<0.01). Conclusion: The occurrence and development of CRC has nothing to do with the expression frequency of CD123+pDCs and is related to the amount of Foxp3+Tregs infiltration in the cancer tissue,The change of DCs subgroups in tumor tissue has a positive relationship with Foxp3+Tregs, which promotes the occurrence and development of CRC.
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Evaluation on clinical effect of Chinese medicine gallnut and andrographolide extract in treatment of periodontal-endodontic combined lesions
ZHAO Zhulan, ZHONG Yong, HUANG Yu, XU Lishuo, LIU Chenguang, ZHANG Li, MA Ning
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  839-843.  DOI: 10.13481/j.1671-587x.20200428
Abstract ( 7 )   PDF (2243KB) ( 8 )  
Objective: To study the clinical effect of Chinese medicine gallnut and andrographolide extract combined with ultrasonic irrigation and periodontal endoscope in the treatment of periodontal-endodontic combined lesions, and to provide a theoretical basis for the effective treatment of periodontal-endodontic combined lesions. Methods: A total of 108 patients with periodontal-endodontic combined lesions were divided into control group, hydrogen peroxide (H2O2) group and experimental group, there were 36 cases in each group. Root canal preparation was carried out in all 3 groups, and distilled water, 3%hydrogen peroxide, gallnut and andrographolide extract were used for root canal irrigation. After preparation, ultrasonic irrigation and different washing agents were used for alternating chemical washing.The medicine was sealed. After the inflammation was eliminated, the root canal was closed. And the X-ray examination results showed that the root canal was filled exactly. After supragingival scaling of the affected tooth, subgingival scaling was performed assisted with periodontal endoscope, the periodontal pockets of the patients in three groups were washed respectively with different washing agents. All patients in various groups were treated for 3 months. The periodontal indexes including modified sulcus bleeding index (mSBI), modified plaque index (mPLI),probing depth(PD)and attachment loss(AL) of the patients in various groups were observed before and after treatment; and the levels of interleukin-1β(IL-1β)in the gingival crevicular fluid(GCF)of the patients in various groups were detected after one course of treatment. Results: There were no significant differences in the periodontal indexes and the levels of IL-1β in GCF of the patients before treatment between various groups(P>0.05).Compared with control group, the PD and AL, the level of IL-1β in GCF of the patients in H2O2 group after treatment were reduced(P<0.05);the mSBI, mPLI, PD and AL, the level of IL-1β in GCF of the patients in experimental group were significantly reduced(P<0.05).Compared with H2O2 group, the mSBI, mPLI, PD and AL, the level of IL-1β in GCF of the patients in experimental group after treatment were obviously reduced(P<0.05). Conclusion: Gallnut and andrographolide exact combined with ultrasonic irrigation and periodontal endoscope has a better therapeutic effect on the periodontal-endodontic combined lesions, which could improve the periodontal indexes and reduce the level of IL-1β in GCF of the patients.
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Expressions of miR-216a-5p and WASL in endometrial carcinoma tissue and their molecular mechanisms of regulating proliferation, migration and invasion of endometrial carcinoma cells
QIN Qiaohong, ZHANG Nan, ZHAO Shujun, LI Hongyu
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  844-850.  DOI: 10.13481/j.1671-587x.20200429
Abstract ( 1 )   PDF (2031KB) ( 10 )  
Objective: To investigate the effects of miR-216a-5p targeting Wiskott-Aldrich syndrome like (WASL) protein on the proliferation, migration and invasion of endometrial carcinoma cells, and to clarify the targeting relationship between miR-216a-5p and WASL gene and its mechanism. Methods: A total of 47 cases of endometrial cancer tissue and normal tissue adjacent to the cancer surgically removed were collected. The miR-216a-5p group,minics negative control(miR-con group) si-con (si-con group), si-WASL (si-WASL group), anti-miR-con (anti-miR-con group), anti-miR-216a-5p (anti-miR-216a-5p group), miR-216a-5p+pcDNA (miR-216a-5p+pcDNA group) and miR-216a-5p+pcDNA-WASL (miR-216a-5p+pcDNA-WASL group) were transfected into the endometrial carcinoma HEC-1-B cells. The expression levels of miR-216a-5p and WASL mRNA and protein in the endometrial carcinoma tissue the adjacent tissue and the HEC-1-B cells in various groups were detected by RT-qPCR and Western blotting methods. MTT assay was used to detect the proliferation activities of the cells in various groups. Transwell assay was used to detect the number of migrated and invasive cells in various groups. The dual luciferase reporter gene assay was used to detect the double luciferase activities in various groups and to determine whether WASL was the target gene of miR-216a-5p. Results: Compared with the adjacent tissue, the expression level of miR-216a-5p in the endometrial carcinoma tissue was significantly decreased (P<0.05), the expression level of WASL mRNA and protein were significantly increased (P<0.05),and they had negative relationship (r=-0.317, P<0.01). Compared with miR-con group and si-con group, the expression level of WASL protein (P<0.01),the proliferation activity of cells (P<0.05) and the number of migrated and invasive cells(P<0.05) in miR-216a-5p group and si-WASL group were significantly decreased. The results of double luciferase reporter gene assay and Western blotting method showed that WASL was the target gene of miR-216a-5p. Compared with miR-216a-5p + pcDNA group, the proliferation activity of cells and the number of migrated and invasive cells in miR-216a-5p + pcDNA-WASL group were significantly increased (P<0.05). Conclusion: The expression level of miR-216a-5p in endometrial carcinoma tissue is decreased. MiR-216a-5p may inhibit the proliferation, migration and invasion of endometrial carcinoma cells through WASL gene, which may be a potential target for the treatment of endometrial carcinoma.
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Preventive effect of early intervention on occurrence of other allergic diseases in infants with food allergy
CHEN ANG, WANG Guilan, LIU Xiangteng, ZHANG Quanshan
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  851-857.  DOI: 10.13481/j.1671-587x.20200430
Abstract ( 3 )   PDF (1184KB) ( 5 )  
Objective: To investigate the preventive effect of early intervention on the occurrence of other allergic diseases in the infants with food allergy, and to provide the reference for exploring the measures to prevent the occurrence of other allergic diseases in the infants with food allergy. Methods: A prospective study design was adopted in our study and 5 712 neonates born in our hospital from January 2017 to October 2017 were selected as the subjects. As of October 2018, a total of 274 infants with food allergy were diagnosed. They were divided into standard intervention group(n=134),standard intervention with probiotics addition group(n=69) and non-standard or non-intervention group(n=71) according to whether standard intervention was carried out and whether probiotics were added along with the intervention. Meanwhile, 187 infants without food allergies at the same period were selected as control group. The incidence of allergic diseases was recorded at 6 and 12 months after enrollment, and the percentages of eosinophil (EOS) and the transforming growth factor-β1 (TGF-β1) levels in peripheral blood of the subjects in various groups were detected. Results: There were no significant differences in the age, gender and family environment among the four groups (P> 0.05).After 6 and 12 months of follow-up, the incidence of eczema, wheezing or persistent cough and asthma had significant differences among the four groups(P<0.01), and the incidence of the above allergic diseases in non-standard or non-intervention group was significantly higher than that in control group (P<0.016 7). After 6 months of follow-up, there was no significant difference in the incidence of allergic rhinitis among the four groups (P>0.05); but after 12 months of follow-up, the incidence of allergic rhinitis in non-standard or non-intervention group was significantly higher than that in control group (P<0.016 7).After 12 months of follow-up, the differences in EOS percentage and TGF-β1 levels in peripheral blood were significantly different among the four groups(P<0.05). Compared with control group,the peripheral blood EOS percentage and the TGF-β1 level in non-standard or non-intervention group were significantly increased (P<0.05); compared with non-standard or non-intervention group, the EOS percentages and the TGF-β1 levels in peripheral blood in standard intervention group and standard intervention with probiotics addition group were decreased (P<0.05). Conclusion: Early intervention in the infants with food allergy can reduce the incidence of allergic diseases. Therefore, comprehensive prevention and treatment measures including the addition of probiotics should be adopted to block the process of allergic diseases in the infants with food allergy.
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Clinical medicine
Follicular dendritic cell sarcoma of small intestine and ileocecal region: A case report and literature review
ZHOU Dongkui, LU Mingqian, FENG Xuesong, LIU Yufei, SONG Hao, XU Liang
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  858-862.  DOI: 10.13481/j.1671-587x.20200431
Abstract ( 11 )   PDF (2270KB) ( 6 )  
Objective: To analyze the clinical features and diagnosis of the patient with follicular dendritic cell sarcoma(FDCS)in the small intestine and ileocecal region, and to improve the clinicians' awareness of the disease. Methods: The clinical materials of a patient with FDCS in the small intestine and ileocecal region were collected,and the clinical features, diagnosis and treatment were analyzed in combination with the relevant literatures. Results: A 61-year-old woman was admitted to hospital because of dull pain in the lower abdomen for 9 d and discovery of pelvic mass for 5 d.The result of color Doppler of uterus and appendages showed the low echo package in the right side of pelvic cavity, and malignant tumor was highly likely. The abdominal enhancement CT results showed the right lower abdomen-pelvic neoplasm accompanying with implantation metastasis around, and the lymph nodes were enlarged. The results of uterus and attachment enhanced MRI of ulterus and appendages showed the right lower abdomen-pelvic lumpsa and the ileocecal region nodules, and malignant tumor was considered. The patient underwent surgical treatment to remove the tumor completely, and the postoperative pathological diagnosis was FDCS in the small intestine and ileocecal region. The patient did not receive radiotherapy or chemotherapy after operation, and then improved and discharged. Conclusion: FDCS is a rare clinical malignant tumor without specific imaging findings. Surgical resection is the first choice of treatment. The pathological diagnosis and immunohistonchemistry are the golden standard for the diagnosis of FDCS. The FDCS patients should be given regular follow-up after operation.
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Modified anterior lacrimal recess approach under nasal endoscope in treatment of primary malignant melanoma of lacrimal sac: A case report and literature review
PEI Ying, ZOU Ying, BA Xiumin, LI Xu
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  863-866.  DOI: 10.13481/j.1671-587x.20200432
Abstract ( 7 )   PDF (1844KB) ( 14 )  
Objective: To analyze the clinical effect of the modified anterior method in the treatment of the patient with primary malignant melanoma of lacrimal sac, and to improve the clinician's understanding of the disease and operation method. Methods: The clinical materials, imaging founding and pathological data of one patient with primary malignant melanoma were collected, the advantages of the modified anterior lacrimal recess approach in the treatment of the disease were analyzed, and the clinical diagnosis criteria and treatment methods were summarized combined with the relevant literature review. Results: The female patient,aged 53 years old, had tears in her right eye for 2 years and tumor in her inner canthus for 6 months and was admitted to the hospital because of tumor of dacrycyst. The tumor was found in inner canthus of the right eye of the patients, which was about 1.7 cm×1.2 cm×1.0 cm in size, with tough texture, unclear boundary, tenderness, and close adhesion with the surrounding skin; when the lacrimal sac area was squeezed, a little blood secretion overflowed from the lacrimal spot. The lacrimal passage was washed and the flushing fluid was flowed back in the original lacrimal point. The orbital CT results showed high density shadow in the right lacrimal sac, high density shadows in the right lacrimal sac area and nasolacrimal duct, and obvious enlargement of the right lacrimal sac. The primary diagnosis was right lacrimal sac and nasolacrimal duct space occupying lesions. Modified anterior lacrimal recess approach under nasal endoscope was used to remove the tumor completely. The postoperative pathology showed that the malignant melanoma involved in the surrounding muscle tissue. Conclusion: The modified anterior lacrimal recess approach under nasal endoscope has the advantages of good field exposure, minimal invasive surgery, easy to operate and low risk.
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Imageology
Analysis on correlation between image features of chest MSCT and prognosis in patients with novel coronavirus pneumonia
GUO Fei, ZHU Lin, XU Hong, XIE Zongyu, ZHANG Li, DENG Xuefei
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  867-874.  DOI: 10.13481/j.1671-587x.20200433
Abstract ( 12 )   PDF (2613KB) ( 8 )  
Objective: To investigate the imaging features of multi-slice spiral computed tomography (MSCT) in the novel coronavirus pneumonia (COVID-19) patients with different prognosis, and to clarify the influencing factors of the prognosis of COVID-19. Methods: The clinical data and the chest MSCT volume scan data of 79 COVID-19 patients, who were admitted in the First Affiliated Hospital of Bengbu Medical College from January 22 to March 8, 2020, were retrospectively analyzed. According to whether there was extensive fibrosis on the MSCT images at discharge or death, the patients were divided into poor prognosis group and well prognosis group. The differences in the MSCT features, the general data and the clinical and laboratory indexes of the patients with different prognosis were compared. Results: There were 74 cases cured in 79 patients, and 5 cases died and the morbidity was 6.3%. The MSCT features at discharge or death were dissipation or ground glass shadow (27 cases, 34.2%), mixed shadow (41 cases, 51.9%) and extensive fibrosis (11 cases, 13.9%).The results of single factor analysis showed that compared with well prognosis group, the number of involved segments of the lung of the patients in poor prognosis group was increased (P<0.05), the temperature was increased(P<0.05), and the percentages of patients with diffuse lesions,the severe patients and the patients with increased white blood cells were increased(P<0.05). The sensitivity and specificity of the number of involved segments for predicting poor prognosis were 72.7% and 80.9%, respectively, while the cut-off value was 9.5. The sensitivity and specificity of temperature for predicting poor prognosis were 54.5% and 92.6%, respectively, while the cut-off value was 39.1℃.The results of Logistic regression analysis showed that the number of involved segments and temperature were the independent risk factors for the poor prognosis of the COVID-19 patients. Conclusion: The number of involved segments in the MSCT images at admission and the temperature are closely related to the prognosis of COVID-19 patients. The number of involved segments more than 9 and the temperature higher than 39.1℃ indicate that the patients are prone to extensive pulmonary fibrosis and poor prognosis.
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Values of 3.0T MRI PDFF and IP-OP in quantificative evaluation on fatty liver
YANG Yiming, LIU Yupin, ZHOU Dongjing, CHEN Zhiyuan, XIAO Huanming, CHI Xiaoling
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  875-880.  DOI: 10.13481/j.1671-587x.20200434
Abstract ( 9 )   PDF (1649KB) ( 9 )  
Objective: To explore the application values of 3.0T magnetic resonance imaging(MRI) proton density fat-fraction(PDFF) and in-phase and opposed-phase(IP-OP) in the quantitative evaluation of liver fat content,and to provide the reference for non-invasive quantitative analysis of fatty liver. Methods: A total of 34 mild fatty liver patients, 29 moderate to severe fatty liver patients and 20 non-fatty liver patients (control group) were selected.All patients were diagnosed by liver biopsy. The hepatic fat fractions(HFF) of patients were detected by MRI-PDFF and MRI-IP-OP techniques. Kruskal-Wallis test was used to compare the differences in HFF between various groups. Mann-Whitney test and Bonferroni correction were used to compare the differences of HFF between two groups. The receiver operating characteristic(ROC) curve was drawn to analyze the diagnossis efficiency of each technique to evaluate the liver steatosis. Results: There were significant differences in HFF of MRI-PDFF and MRI-IP-OP of the patients between various groups(P<0.01).The HFF of MRI-PDFF of the patients in moderate and severe fatty liver group was higher than those in control group and mild fatty liver group(P<0.01), and the HFF of the patients in mild fatty liver group was higher than that in control group(P<0.01).The HFF of MRI-IP-OP the patients in moderate and severe fatty liver group was higher than those in control group and mild fatty liver group(P<0.01), and the HFF of the patients in mild fatty liver group was higher than that in control group (P<0.05).The ROC curve showed that the areas under curve(AUC) of MRI-PDFF and MRI-IP-OP in the diagnosis of mild fatty liver were 0.931 and 0.874, respectively, and the AUC of MRI-PDFF and MRI-IP-OP in the diagnosis of moderate and severe farry liver were 0.928 and 0.913, respectively. Conclusion: For the mild and moderate to severe fatty liver, MRI-PDFF has a more higher diagnosis efficiency,and it is a accurate, reliable,and non-invasive quantitative analysis method for liver steatosis.
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Research progress in regulation of proliferation, differentiation and glucose and lipid metabolism of 3T3-L1 adipocytes by traditional Chinese medicine
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  881-887.  DOI: 10.13481/j.1671-587x.20200435
Abstract ( 6 )   PDF (2073KB) ( 13 )  
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Research progress in prevention and treatment of intraoperative and postoperative lymphatic cysts in female patients with malignant tumor pelvic
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  888-893.  DOI: 10.13481/j.1671-587x.20200436
Abstract ( 18 )   PDF (1482KB) ( 17 )  
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Research progress in relationship between GRIM-19 and occurrence and development of urogenital reproductive neoplasms
Journal of Jilin University(Medicine Edition). 2020, 46 (04):  894-898.  DOI: 10.13481/j.1671-587x.20200437
Abstract ( 12 )   PDF (1481KB) ( 10 )  
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