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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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Table of Content
28 January 2021, Volume 47 Issue 1
Research in basic medicine
Effect of low dose ionizing radiation on hippocampal neurons in STZ-induced diabetic rats
Fei LIU,Shuo LIANG,Yuexuan WANG,Lijing QIN,Wei GUO,Zhicheng WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  1-7.  DOI: 10.13481/j.1671-587x.20210101
Abstract ( 341 )   HTML ( 2 )   PDF (895KB) ( 142 )  
Objective

To investigate the effects of low dose ionizing radiation (LDR) on the cell cycle, calcium ion concentration([Ca2+]i) and mitochondrial membrane potential(MMP) of hippocampal neurons in the diabetic rats, and to elucidate the protective mechanisms of LDR on nerve injury.

Methods

The Wistar rat models of diabetes mellitus (DM) were established with streptococci (STZ).Sixty rats were divided into normal control group, DM X-ray irradiation group, DM+25 mGy X-ray irradiation group, DM+50 mGy X-ray irradiation group and DM+75 mGy X-ray irradiation group; there were 12 rats in each group. Except normal control and DM groups, the rats in other groups were administered with 25, 50 and 75 mGy LDR every other day for 12 times, and the body weights of the rats were monitored. After 4 weeks of feeding, the rats were sacrificed after anesthesia. The hippocumpus tissue of the rats in various groups was removed and single cell suspension was prepared. The percentages of hippocampal neurons at different cell cycles, ([Ca2+]i) and MMP were detected by flow cytometry, respectively.

Results

With the prolongation of time,the body weight of the rats in normal control group was increased gradually. Compared with normal control group, the body weights of diabetic rats in DM group, DM+25 mGy X-ray irradiation group, DM+50 mGy X-ray irradiation group and DM+75 mGy X-ray irradiation group were significantly decreased (P<0.05); compared with DM group, the body weights in DM+50 mGy group and DM+75 group mGy group were significantly increased(P<0.05). Compared with normal control group, the percentages of hippocampal neurons in S phase and [Ca2+]i of the rats in DM group were increased significantly(P<0.05), while MMP was decreased significantly (P<0.05); compared with DM group, the percentages of hippocampal neurons in S phase and [Ca2+]i of the rats in DM+50 mGy X-ray irradiation group and DM+75 mGy X-ray irradiation group were decreased significantly (P<0.05), while MMP of hippocampal neurons was increased significantly (P<0.05); compared with DM + 25 mGy X-ray irradiation group, the [Ca2+]i in hippocampal neurons in DM+75 mGy X-ray irratiaiton group was decreased significantly(P<0.05), while MMP of hippocampal neurons in DM+50 Gy X-ray irradiation group and DM+75 Gy X-ray irradiation group were increased significantly (P<0.05).

Conclusion

LDR can improve the S phase delay, the increase of [Ca2+]i and the decrease of MMP induced by DM of the hippocampal neurons of the rats to play the neuroprotective roles.

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Synergistic inhibitory effect of aaptamine and cisplatin on cisplatin resistant human lung adenocarcinoma A549/DDP cells
Shuang MIAO,Na NI,Lijuan YANG,Yan WU,Xuelin LI,Hongliang DONG,Kaikai GONG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  8-15.  DOI: 10.13481/j.1671-587x.20210102
Abstract ( 495 )   HTML ( 0 )   PDF (1475KB) ( 110 )  
Objective

To investigate the synergetic inhibitory effect of aaptamine and cisplatin (DDP) on the DDP resistant human lung cancer A549/DDP cells, and to clarify its possible molecular mechanism.

Methods

The A549/DDP cells in logarithmic phase were selected and cultured with different concentrations of aaptamine and DDP for 48 h; the medium inhibitory concentrations(IC50) of aaptamine and DDP were detected by CCK-8 assay; the combination index (CI) of aaptamine and DDP were analyzed by Chou-Talalay method and the optimal combination concentrations were confirmed. The cells were divided into control group, aaptamine group(5 mg·L-1), DDP group(1 mg·L-1), and combination group (5 mg·L-1 aaptamine +1 mg·L-1 DDP). The proliferation activities of cells in various groups were detected by CCK-8 assay.The number of clones of the cells in various groups was determined by clonal formation assay. The migration abilities of the cells in various groups were detected by scratch assay. The apoptotic rates of cells in various groups were detected by double-staining flow cytometry. Western blotting method was performed to detect the expression levels of resistance-related proteins in the cells in various groups.

Results

The IC50 values of aaptamine and DDP for the A549/DDP cells were 19.45 mg·L-1 and 12.86 mg·L-1, respectively. Co-treatment with aaptamine and DDP produced a synergistic effect at any selected concentrations in the A549/DDP cells. The proliferation assay results showed that compared with control group, aaptamine group and DDP group, the proliferation activity of the cells in combination group was decreased (P<0.01). The clone formation assay results showed that compared with control group, aaptamine group and DDP group, the number of clones in combination group was decreased (P<0.05 or P<0.01). The scratch assay results showed that compared with control group, aaptamine group and DDP group, the migration rate of cells in combination group was decreased significantly (P<0.01). The flow cytometry results showed that compared with control group, aaptamine group and DDP group, the apoptotic rate of A549/DDP cells in combination group was markedly increased (P<0.05 or P<0.01). The Western blotting results showed that compared with control group, aaptamine group and DDP group, the expression level of resistance related proteins ATP binding cassette subfamily G member 2(ABCG2) in the cells in combination group was decreased (P<0.05 or P<0.01),and the ratio of Bax/Bcl-2 was increased (P<0.05 or P<0.01).Compared with control group,the expression level of excision repair cross complementing group 1(ERCC1) protein in combination group was decreased (P<0.05).

Conclusion

Aaptamine has a synergistic inhibitory effect with DDP in the A549/DDP cells and its mechanism may be related to inhibiting proliferation, inducing apoptosis and decreasing the expression levels of resistance-related proteins ABCG2 and ERCC1.

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Construction of human hepatocyte growth factor gene lentivirus vector and its expression in bone marrow mesenchymal stem cells
Chongjun XU,Hefan HE,Qun LIN,Tao ZHANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  16-24.  DOI: 10.13481/j.1671-587x.20210103
Abstract ( 409 )   PDF (1542KB) ( 99 )  
Objective

To construct the human hepatocyte growth factor (HGF) gene lentivirus vector pNL-HGF-EGFP and transfect the bone marrow mesenchymal stem cells(BMSCs),and to detect the expression of HGF gene in the BMSCs.

Methods

The recombinant lentivirus plasmid pNL-HGF-EGFP was constructed by using restriction enzyme digestion to obtain HGF gene sequence and cloned into the lentivirus vector pNL-EGFP.The lentivirus vector plasmid pNL-EGFP and recombinant lentivirus plasmid pNL-HGF-EGFP were co-transfected into the 293T cells with lentivirus assisted packaging plasmid, respectively. The lentivirus was packaged and the titer of lentivirus was determined.The BMSCs infected with pNL-EGFP were used as control group, and the BMSCs infected with pNL-HGF-EGFP were used as experimental group.RT-PCR and ELISA methods were used to detect the expression levels of HGF mRNA and the levels of HEG protein in the BMSCs in control group and experimental group.

Results

The electrophoresis and sequencing results showed that the target gene sequence of the recombinant lentivirus plasmid was consistent with the HGF gene sequence published in GenBank.The packaged lentivirus titer in control group was 1.2×107 TU·mL-1, and the lentivirus titer in experimental group was 1.5×106 TU·mL-1.The RT-PCR detection results showed that the expression level of HGF mRNA in the BMSCs in experimental group was higher than that in control group(P<0.05).The ELIAS results showed that the level of HGF protein in the BMSCs in experimental group was significantly higher than that in control group (P<0.05).

Conclusion

The lentiviral vector pNL-HGF-EGFP is successfully constructed, and the expression level of HGF mRNA and and the level of HGF protein in the BMSCs are significantly increased.

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Inhibitory effect of lithium chloride on Aβ1-42-induced astrocyte glutamate release and its protective effect on hippocampal neuronal injury
Qi LIU,Xianchen ZHANG,Xiangyu LI,Yumiao LIN,Yanqi YANG,Enzhi YAN
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  35-43.  DOI: 10.13481/j.1671-587x.20210105
Abstract ( 357 )   PDF (1768KB) ( 159 )  
Objective

To investigate the inhibitory effect of lithium chloride on amyloid β1-42(Aβ1-42)-induced astrocyte glutamate release, and to explore its protective effect on hippocampal neuronal injury and mechanism.

Methods

The astrocytes(AS) and hippocampal neurons were primarily cultured.The AS were divided into control group and Aβ1-42+different doses (0.25,0.50 and 1.00 mmol?L-1)of lithium chloride groups. After treated with lithium for 2 weeks, the AS were stimulated with 250 nmol?L-1 1-42.The Ca2+ level in AS was detected by fluorescence probe technique, the release amount of glutamate of AS was measured by HPLC method, and Western blotting method was performed to detect the expression levels of transient receptor potential channel 1(TRPC1),Na+/Ca2+ exchanger 1(NCX1) and CACNA1C (Cav1.2) proteins in AS. The cultured hippocampal neurons were divided into control group, Aβ1-42 group and Aβ1-42+different doses of lithium chloride groups. The conditioned medium containing AS or not treated with Aβ1-42 were added,respectively.The total dendrite branch length (TDBL), number of primary-order dendrite(PDN), maximum branch order(MBO) of hippocampal neurons were observed by inverted phase-contrast microscope.The amount of NO release in culture solution was measured by Griess method.

Results

Compared with control group, the levels of Ca2+ in AS in Aβ1-42 +different doses of lithium chloride groups were significantly decreased (P<0.05), the Ca2+ influx was reduced(P<0.05),the release amounts of glutamate were decreased significantly (P<0.05), and the expression levels of TRPC1 protein were decreased significantly (P<0.05), but the expression levels of NCX1 and Cav1.2 proteins had no significant differences (P>0.05). Compared with control group, the release amount of NO in hippocampal neurons in Aβ1-42 group was significantly increased(P<0.05), and TDBL, PDN and MBO were significantly decreased(P<0.01). Compared with Aβ1-42 group, the release amounts of NO in hippocampal neurons in Aβ1-42 +different doses of lithium chloride groups were decreased (P<0.01), and TDBL, PDN and MBO were significantly increased(P<0.01). The release amounts of NO in hippocampal newrons in various groups had significant difference after cultured with Aβ1-42-treated culture medium without AS(P>0.05).

Conclusion

Lithium chloride has inhibitory effect on Aβ1-42-induced Ca2+-dependent glutamate release in AS,and its mechanism may be related to down-regulation of TRPC1 receptor expression;lithium chloride can alleviate the injury of hippocampal neurons by inhibiting the release of NO in the neurons.

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Activity and stability of reductase of Mesonia sp. S52 Cr(Ⅵ) and its optimization of Cr(Ⅵ) reduction conditions
Jiayao LI,Xiaoxia LI,Qiuying AN,Chun FAN,Dongbei GUO,Chen TANG,Min ZHANG,Xieryazdan SANGDUHAX,Ran ZHAO
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  44-52.  DOI: 10.13481/j.1671-587x.20210106
Abstract ( 335 )   HTML ( 0 )   PDF (1075KB) ( 43 )  
Objective

To explore the effects of temperature and pH value on the enzyme activity and stability of Mesonia sp. S52 Cr(Ⅵ) and optimize the reduction conditions, and to study the effects of metal ions and small molecules on the reducing ability of the reductase.

Methods

The seed solution of the S52 strain was inoculated and cultured overnight in LB medium. The extracellular active substance obtained by centrifugal filtration of the bacterial solution, the intracellular active substance obtained by ultrasonic ice-breaking bacteria and the S52 whole bacterial solution were added to the LB medium containing 50 mg·L-1 Cr(Ⅵ) and cultured at 37 ℃, pH 8.0. Benzyl dihydrazide spectrophotometry was used to determine the Cr (Ⅵ) concentrations in the solution at 0, 3, 6,12, 24 and 48 h, and then the reduction rates of Cr(Ⅵ), the relative activities and relative stabilities of intracellular and extracellular enzymes were calculated. Using a multi-factor mixed experiment design, the temperatures were set as 25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃, 50 ℃, and the pH values were 5.0, 6.0, 7.0, 8.0, 9.0, 10.0; the reduction rates of Cr(Ⅵ) by reductases at 0, 3, 6 and 12 h were calculated, then the effects of different temperatures and pH values on the reduction rate of reductase were analyzed, and the optimal reduction conditions were optimized. 0.2 mmol·L-1 Cu2+, Mn2+ and Cd2+ solution, 1mmol·L-1 SDS and EDTA, 1% Triton X-100, Tween 80 were added to the medium separately as treatment groups, and the untreated culture medium was used as control group. They were reacted with 50 mg·L-1 Cr(Ⅵ) for 3 h under the optimal reduction conditions. The reduction rates of Cr(Ⅵ) were measured and calculated, and the changes of reduction rates in different metal ions and small molecules treatment groups were compared.

Results

At 3 h, the reduction rates of Cr(Ⅵ) by intracellular and extracellular enzymes reached the highest, and the reduction ability of intracellular enzymes was higher than that of extracellular enzymes (P<0.05). When the temperature was constant and the pH value was from 5.0 to 6.0, the reduction rates of Cr(Ⅵ) by intracellular and extracellular enzymes were increased significantly; when the pH values were gradually increased, the reduction rates of Cr(Ⅵ) were decreased significantly. Under certain pH condition, with the increase of temperature, the intracellular enzyme reduction rates were gradually increased, and the extracellular enzyme reduction rates were decreased. The differences in reduction rates between different temperature and pH groups were statistically significant (P<0.05), and the interaction between temperature and pH was significant (P<0.05). Compared with control group, the enzymatic reduction rate in Cu2+ treatment group reached to 46.4% (P<0.05),the reduction rates in Cd2+ and Mn2+ treatment groups were decreased(P<0.05); there was no statistically significant difference in the reduction rate of Cr(Ⅵ) between small molecular substance Triton X-100 and control group (P>0.05), while the reduction rates of Cr(Ⅵ) in Tween 80, EDTA and SDS treatment groups were lower than that in control group(P<0.05).

Conclusion

The Cr(Ⅵ) reductase of strain S52 can play a reducing role both inside and outside the cells, and the reducing effect of intracellular enzyme is stronger than that of extracellular enzyme. The optimal temperature of intracellular enzymes is 35 ℃-45 ℃, and the optimal pH value is 7.0; the optimal temperature of extracellular enzymes is 25 ℃-30 ℃, and the optimal value is 7.0. Different metal ions and small molecules have different effects on the activities of reductases.

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Improvement effect of curcumin on cerebral microcirculation disorder in hypercholesterolemia model rats and its mechanism
Hongfang LI,Shao YANG,Mohan CAO,Lili YUAN,Xiaojin LI,Caixing SHI,Zhuoya WANG,Bailiu YA
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  53-58.  DOI: 10.13481/j.1671-587x.20210107
Abstract ( 450 )   HTML ( 0 )   PDF (781KB) ( 507 )  
Objective

To study the effects of curcumin on the oxidative stress and inflammation status of cerebral microcirculation in the hypercholesterolemia model rats induced by high cholesterol diet,and to elucidate their mechanisms.

Methods

Thirsty SD rats were randomly divided into control group, model group and curcumin group(n=10). The rats in control group were given ordinary diet and the rats in other two groups were given high cholesterol diet daily for 28 d.At the same time, the rats in curcumin group were given 200 mg·kg-1·d-1 curcumin by gavage, and the rats in control group and model group were given 1% sodium carboxymethylcellulose solution; once a day for 28 d. The sera of rats were taken and the levels of blood lipids were detected by enzyme method. The cerebral microvessels of rats were extracted; the activity of superoxide dismutase (SOD)in cerebral microvessel was detected by xanthine oxidase method; the level of malondialdehyde (MDA) was detected by thiobarbituric acid method; the expression levels of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1(VCAM-1) and E-selectin proteins in the cerebral microvessel endothelial cells of the rats were detected by Western blotting method.

Results

Compared with control group, the total cholesterol(TC),triglyceride(TG),and low density lipoprotein-cholesterol( LDL-C) in serum of the rats in model group were significantly increased(P<0.01),the high density lipoprotein-cholesterol(HDL-C) in serum was significantly decreased (P<0.01),the SOD activity in cerebral microvessel was significantly decreased (P<0.01), the level of MDA was significantly increased(P<0.05), and the expression levels of ICAM-1,VCAM-1 and E-selectin proteins in cerebral microvessel endothelial cells were significantly increased (P<0.05 or P<0.01). Compared with model group, the TC, TG, LDL-C in serum of the rats in curcumin group were significantly decreased (P<0.05), the HDL-C in serum was significantly increased (P<0.05), the SOD activity in cerebral microvessel was significantly increased (P<0.05), the level of MDA was significantly decreased (P<0.05), and the expression levels of ICAM-1, VCAM-1 and E-selectin proteins in cerebral microvessel endothelial cells were significantly decreased(P<0.05).

Conclusion

Curcumin may ameliorate the cerebral microcirculation disorder by improving the blood lipid status of the hypercholesterolemia rats, increasing the ability of antioxidant enzyme, reducing oxidative stress damage and decreasing the expressions of adhesion factors in the endothelial cells of cerebral microcirculation.

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Regulatory effects of syndecan-1 on migration, invasion and cell cycle of tongue squamous cell carcinoma CAL27 cells
Lu LIU,Tianfu ZHANG,Xiaofeng WANG,Chenfei KONG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  59-65.  DOI: 10.13481/j.1671-587x.20210108
Abstract ( 279 )   PDF (1388KB) ( 95 )  
Objective

To study the effect of overexpression of syndecan-1 (SDC1) on migration, invasion, cell cycle, and intracellular reactive oxygen(ROS) level of the tongue squamous cell carcinoma CAL27 cells, and to elucidate the potential mechanism of SDC1 in the occurrence and development of tongue squamous cell carcinoma.

Methods

The tongue squamous cell carcinoma CAL27 cells stably high-expressing ptt5-SDC1 established successfully in the previous experiment were used as experimental group, and the CAL27 cells stably transfected with ptt5 empty were used as control group. Western blotting method was used to detect the expression levels of SDC1 protein in the cells in two groups. Cell scratch test was used to detect the healing rates of cell scratch in two groups; Transwell chamber assay was performed to detect the number of migration and invasion cells;flow cytometry was used to detect the percentage of CAL27 cells in different cell cycles and the levels of intracellular ROS.

Results

Compared with control group, the expression level of SDC1 protein in CAL27 cells in experimental group was significantly increased(P<0.05), the healing rate of scratch of the cells was significantly reduced (P<0.05), the number of migration and invasion cells was significantly reduced (P<0.01), the percentage of cells in G0/G1 phase was significantly increased (P<0.05), and the inrracellular ROS level was significantly increased (P<0.05).

Conclusion

SDC1 overexpression can inhibit the invasion and metastasis of tongue squamous cell carcinoma CAL27 cells, and its mechanism may be related to cell cycle arrest and the pathway mediating apoptosis.

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Screening and identification of target proteins interacting with GINS2 in human promyelocytic leukemia cDNA library
Xi ZHANG,Yuqing PAN,Xin YU,Yating ZHENG,Chengbin GUO,Na XU,Liangxiao WANG,Liang HE,Li YANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  66-72.  DOI: 10.13481/j.1671-587x.20210109
Abstract ( 349 )   HTML ( 0 )   PDF (1096KB) ( 78 )  

Objective: To screen the target proteins interacting with human GINS2(go-ichi-ni-san) by yeast two hybrid system, and to elucidate its pathogenesis in acute promyelocytic leukemia (APL) and related mechanism.

Methods

The constructed bait plasmid pGBKT7-GINS2 and human promyelocyte cDNA library plasmids were simultaneously transformed into the yeast AH109 cells by heat shock method to select the library proteins interacting with GINS2 coding proteins. Then, multiple reporter positive clones detection and DNA sequencing were used for bioinformatics analysis of the genes encoding the interacting proteins.

Results

A total of 15 initial positive clones were successfully screened from the yeast two hybrid library, among which 10 clones could activate ADE2 and HIS3 reporter genes,and 13 clones could activate LacZ reporter gene. After DNA sequencing and BLAST analysis of 10 positive clones which could grow in SD/-Trp/-Leu/-His/-Ade defective culture medium were performed, except for the sequencing failure of No. 8 and No. 9, the remaining 8 positive clones belonged to the eight different protein-coding genes, namely RNA binding motif protein 39(RBM39), ubiquitin like modifier activating enzyme 1(UBA1),metallothionein 2A (MT2A),mitochondrial calcium uptake 1(MICU1),N-ethylmaleimide sensitive fusion protein cofactor 1(NSFL1C), fatty acyl-CoA reductase 1(FAR1), dual specificity phosphatase 1(DUSP1) and cell surface antigen 14(CD14).

Conclusion

A total of eight functional proteins that interact with GINS2 are successfully screened, and it is speculated that the target proteins may play an important role in cell damage, transcriptional regulation, proliferation and metabolism.

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Inhibitory effect of nitidine chloride on epithelial-mesenchymal transition of glioma cells through JAK2/STAT3 signaling pathway
Mingbo JIA,Ying SUN,Ying WANG,Yanke SONG,Liyan ZHAO
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  73-81.  DOI: 10.13481/j.1671-587x.20210110
Abstract ( 569 )   HTML ( 0 )   PDF (1740KB) ( 50 )  
Objective

To investigate the inhibitory effect of nitidine chloride (NC) on the epithelial-mesenchymal transition (EMT) of glioma cells, and to clarify signaling mechanism of the inhibitory effect.

Methods

The human glioma U87 cells cultured in vitro were divided into control group (without treatment),EMT group [EMT induced by transforming growth factor-β1(TGF-β1)] and different concentrations (2.5, 5.0, 7.5 and 10 μmol·L-1) of NC groups (different concentrations of NC+10 μg·L-1 TGF-β1). After 48 h of treatment, the cell migration rates,the invasion rates and the expression levels of EMT-related proteins and JAK2 / STAT3 pathway-related proteins were detected by wound healing test, Transwell invasion test and Western blotting method, respectively. When inducing EMT in the U87 cells, the JAK2 / STAT3 pathway was blocked by STAT3 inhibitor WP1066. The U87 cells were divided into control group (without treatment), EMT group (10 μg·L-1 TGF-β1) and EMT + WP1066 group (10 μg·L-1 TGF-β1 + 8 μmol·L-1WP1066); the expression levels of EMT-related proteins in the cells in various group were detected by Western blotting method.

Results

The cell migration rate and invasion rate in EMT group were significantly higher than those in control group (P<0.01), while the cell migration rate and invasion rate in NC group were significantly lower than those in EMT group (P<0.01). Compared with control group, the expression level of E-cadherin in the U87 cells in EMT group was decreased(P<0.01), while the expression levels of N-cadherin, vimentin, β-catenin, and the transcription factors Snail, Slug and Twist1 proteins were increased (P<0.05 or P<0.01). Compared with EMT group, the expression levels of E-cadherin in different concentrations of NC groups were increased (P<0.05 or P<0.01), and the expression levels of N-cadherin, vimentin, β-catenin, and transcription factors Snail, Slug and Twist1 proteins were significantly reduced (P<0.01) in a concentration-dependent manner; 7.5 μmol·L-1NC could significantly inhibit the EMT of glioma cells. Compared with control group,the expression levels of JAK2 and p-JAK2 proteins in the cells in EMT group had no significant differences(P>0.05),and the expression levels of STAT3 and p-STAT3 in the cells were significantly decreased(P<0.05).Compared with EMT group,the expression levels of JAK2,p-JAK2, STAT3 and p-STAT3 proteins in the cells in different NC groups were significantly reduced (P<0.01). In JAK2/STAT3 pathway blocking experiment by STAT3 inhibitor WP1066, the expression levels of E-cadherin proteins in the cells in EMT+WP1006 group was significantly increased(P<0.01),and the expression levels of N-cadherin,vimentin,β-catein Twist1,Slug and Snail proeins were significantly decreased(P<0.01).

Conclusion

NC can inhibit the EMT of glioma cells, and the inhibitory effect is related to JAK2/STAT3 signaling pathway.

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Preparation and bone-binding properties of 3D printed titanium alloy implants
Rui WANG,Meihua LI,Wanlin ZHOU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  82-88.  DOI: 10.13481/j.1671-587x.20210111
Abstract ( 564 )   HTML ( 1 )   PDF (1398KB) ( 182 )  
Objective

To observe the osseointegration of 3D printed implants and classic Straumann implants after implanted in the rabbits,and to provide the theoretical basis for its clinical application.

Methods

The Ti-6A1-4V(TC4) implants were made by 3D printed method,and the classic Straumann implants were used as controls.The animal models were established by implanting bilateral femur implants in 6 rabbits; one TC4 implant and one Straumann implant were implanted on each side,and there were 12 T4C implants((TC4 implant group) and 12 Straumann implants (Straumann implant group) in all. At 2, 4 and 8 weeks after the operation, two rabbits in each group were killed respectively, and the tissue around the implants was taken out; toluidine blue staining was used to observe osteoblasts, new bone formation and bone tissue repair, and methylene blue acid fuchsin staining was used to observe the new bone repair and mineralization.The surface structure changes of TC4 implants after operation were observed by scanning electron microscope(SEM),and the element composition of TC4 implants after operation was analyzed by energy dispersive X-ray spectrometer(EDS).

Results

The toluidine blue staining results showed that at 2 weeks after the operation, the deep staining of nucleus and linear arrangement of light blue osteoblasts on the edge of bone tissue close to the implants of the rabbits in two groups were found; at 4 weeks after the operation, the existence of light stained new collagen fiber-like structure around the implants in two groups was seen, and the arrangement was not orderly; at 8 weeks after the operation,the light blue new bone formation with continuous structures was found in one side close to the implants of the rabbits in two groups, containing a large number of bone cells, and the boundary between new bone and original mineralized bone was clear. The methylene blue acid fuchsin staining results showed that at 2 weeks after the operation, the implants in two groups showed active osteoblast aggregation around the implants; at 4 weeks after the operation, new and red stained osteoid structures were found between the implants and the original mineralized bone in two groups; at 8 weeks after the operation, new bone formation with certain degree of calcification was seen on the surface of the implants in two groups, which was dark red,and the degree of calcification in Straumann group was slightly better than that in TC4 group. The SEM results showed that the TC4 implants completed in vivo experiment were basically consistent with the preoperative TC4 implants in surface structure, and no obvious structural defects were found.The results of EDS showed that the TC4 implant completed in vivo experiment contained not only the elements consistent with the preoperative TC4 implants, but also the major elements Ca, Mg and trace elements Si.

Conclusion

The 3D printed TC4 implants have good structural stability and biocompatibility, and can achieve the similar bone binding with Straumann implants invivo.

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Expressions of MMP-3 and CCN2 in dental pulp tissue of rats with dental pulp injury and their significances
Mengjie LI,Jinfang XIE,Shuo YIN,Xia LIU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  89-95.  DOI: 10.13481/j.1671-587x.20210112
Abstract ( 442 )   HTML ( 0 )   PDF (1546KB) ( 147 )  
Objective

To establish dental pulp injury rat models, and to preliminarily illustrate the role of matrix metalloproteinase-3(MMP-3) and connective tissue growth factor 2(CCN2) in promoting the repair of dental pulp tissue after injury.

Methods

A total of 28 rats were divided into control group and 0 h, 6 h, 1 d, 3 d, 7 d and 14 d after injury groups.Except control group,the rat models of pulp injury of maxillary molars were established by simple pulp exposure. The pulp tissue of rats in various groups were obtained at different time after injury. HE and immunohistochemistry staining methods were used to observe the pathomorphology of pulp tissue and the localization and the expression levels of MMP-3 and CCN2 proteins.

Results

The HE staining results showed that the four layers of odontoblasts, depleted cell layers, multicellular layers and pulp proper were clearly visible in the pulp tissue of the rats in control group;the blood vesels were slightly dilated in the pulp tissue of the rats in 0 h injury group; the blood vessels were significantly dilated in the pulp tissue of the rats in 6 h after injury group; the gathered inflammatory cells in the pulp tissue of the rats were found in 3 d after injury group; the new blood vessels, fibroblast proliferation, and new dentin-like cells in the pulp tissue of the rats were seen in 7 d after injury group; reparative dentin formation was seen in 14d after injury group. The immunochemistry results showed that no positive staining of MMP-3 and CCN2 was found in normal pulp of the rats in control group; in 0 h after injury group and 6 h after injury group, the weak expressions of MMP-3 and CCN2 were seen;the positive expressions of MMP-3 and CCN2 were found in odontoblasts and cellular matrix under the injury site in 1 d after injury group; in 3 d and 7 d after injury groups, strongly positive expressions of MMP-3 and CCN2 were found in the odontoblasts, fibroblasts and neutrophils (P<0.05); in 14 d after injury group, the positive expressions of MMP-3 and CCN2 in reparative dentin of the rats were significantly reduced.Compared with control group,the expression levels of MMP-3 and CCN2 proteins in pulp tissue of the rats in different time after injury groups were significantly increased(P<0.05);compared with 0 h after injury group, the expression levels of MMP-3 and CCN2 proteins in pulp tissue of the rats in 6 h after injury group had no significant difference(P>0.05),and the expression levels of MMP-3 and CCN2 proteins in pulp tissue of the rats in 1 d,3 d,7 d and 14 d after injury groups were significantly increased(P<0.05).

Conclusion

The expressions of MMP-3 and CCN2 in the rat injured dental pulp tissue first increase and then decrease, suggesting that MMP3 and CCN2 may promote self-repair after pulp injury.

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Effects of Tanreqing Injection on T lymphocyte subsets in peripheral blood of obstructive sleep apnea-hypopnea syndrome rats
Jing XUE,Zhiming LIU,Yongfeng WU,Kaifeng DONG,Qingfeng WANG,Jinmei ZU,Haijing ZHANG,Bin LOU,Xiangling KONG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  96-101.  DOI: 10.13481/j.1671-587x.20210113
Abstract ( 393 )   HTML ( 0 )   PDF (902KB) ( 81 )  
Objective

To investigate the effects of Tanreqing Injection on the T lymphocyte subset function in peripheral blood of the rats with obstructive sleep apnea hypopnea syndrome(OSAHS), and to illustrate their mechanisms.

Methods

Fifty male SD rats were randomly divided into control group, model group, low, medium and high doses of Tanreqing Injection groups (n =10). The OSAHS rat models were established by alternating cycles of hypoxia(6%-8% O2 in N2 for 40 s) and normoxia (2l% O2 in N2 for 80 s) in model group, low, medium and high doses of Tanreqing Injection groups, 8 h every day. The rats in low, medium and high doses of Tanreqing Injection groups were given 0.9, 1.8 and 3.6 mL·kg-1 Tanreqing Injection by gavage, while the rats in control group and model group were given equal volume of normal saline. The spleen indexes and thymus indexes of the rats in various groups were measured after 14 d of continuous administration. MTT assay was used to detect the proliferation activities of T lymphocytes of the rats in various groups. Flow cytometry was used to detect the percentages of different T lymphocyte subsets in peripheral blood of the rats in various groups. ELASA was used to detect the serum levels of tumor necrosis factor-α (TNF-α)and interleukin- 6(IL-6) of the rats in various groups.

Results

Compared with control group, the spleen index, the thymus index, the proliferation activity of T lymphocytes, the percentage of CD4+ T lymphocytes, the CD4+/CD8+ T lymphocyte ratio of the rats in model group were significantly decreased (P<0.05), while the percentage of CD8+ T lymphocytes, the levels of serumTNF-α and IL-6 were significantly increased (P<0.05). Compared with model group, the spleen indexes, the thymus indexes, the proliferation activities of T lymphocytes, the percentages of CD4+ T lymphocytes, the CD4+/CD8+ T lymphocytes ratios of the rats in low, medium and high doses of Tanreqing Injection groups were significantly increased(P<0.05 ),and the percentage of CD8+ T lymphocytes and the levels of serumTNF-α and IL-6 in the peripheral blood were significantly increased (P<0.05). Compared with low dose of Tanreqing Injection group, the proliferation activity of T lymphocytes in the high dose of Tanreqing Injection group was significantly increased (P<0.05), and the levels of TNF-α and IL-6 in serum of the rats in medium and high doses of Tanreging Injection groups were significantly decreased (P<0.05).

Conclusion

Tanreqing Injection can protect the rats with OSAHS by regulating the balance of T lymphocyte subsets and inhibiting inflammation response.

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Protective effect of crocin on myocardium injury induced by ischemia and hypoxia in rats and its mechanism
Jiancheng HUANG,Shichao GUO,Pujuan LIU,Yanbo DONG,Hongying LI,Ying LYU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  102-109.  DOI: 10.13481/j.1671-587x.20210114
Abstract ( 635 )   HTML ( 0 )   PDF (1147KB) ( 426 )  
Objective

To investigate the protective effect of crocin on the myocardium injury induced by ischemia and hypoxia of the rats, and to analyze its mechanism.

Methods

A totol of 50 SD rats were randomly divided into sham operation group, model group, low dose (20 mg·kg-1) of crocin group, medium dose (40 mg·kg-1)of crocin group and high dose (80 mg·kg-1) of crocin group, with 10 rats in each group. The myocardial ischemia reperfusion injury rat models were established by ligation of left anterior descending coronary artery and reperfusion. The morphology of myocardium tissue of rats in various groups was observed by HE staining. TUNEL was used to detect the apoptosis of myocardial cells, and the apoptotic index (AI) was calculated. The expression levels of B-cell lymphoma-2(Bcl-2), Bcl-2 associated X protein(Bax) and caspase-3 mRNA and proteins in myocardium tissue of the rats were determined by RT-PCR and Western blotting methods. The levels of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), lactate dehydrogenase (LDH), and creatine kinase (CK) in myocardium tissue of the rats in various groups were detected with detection kits.

Results

The myocardial fibers of the rats in sham operation group were arranged neatly; the myocardial fibers of the rats in model group were disordered, edema occurred, and the inflammatory cells were abundant in myocardial stroma;compared with model group, the inflammation and infiltration of the rats in low dose of crocin group were significantly reduced, and the myocardial fiber edema was significantly reduced; there was no significant edema of myocardial fibers with little inflammatory infiltration in medium and high doses of crocin groups. Compared with sham operation group, the AI of myocardium of the rats in model group was increased (P<0.05),the expression levels of Bcl-2 mRNA and protein in myocardium tissue of the rats in model group were significantly decreased (P<0.05), the expression levels of Bax and caspase-3 mRNA and proteins were significantly increased (P<0.05), the serum MDA level was significantly increased (P<0.05),the SOD activity was significantly decreased (P<0.05), and the LDH and CK activities were significantly increased (P<0.05). Compared with model group, the AI of myocardium of the rats in low, medium and high doses of crocin groups were significantly decreased (P<0.05),the expression levels of Bcl-2 mRNA and protein in myocardium tissue of the rats were significantly increased (P<0.05), the expression levels of Bax and caspase-3 mRNA and protein were significantly decreased(P<0.05), the serum MDA levels were significantly decreased (P<0.05), the SOD activities were significantly increased (P<0.05),and the LDH and CK activities were significantly decreased (P<0.05). Compared with low dose of crocin group, the AI of myocardium of the rats in medium and high doses of crocin groups were significantly decreased(P<0.05), the expression levels of Bcl-2 mRNA and protein in myocardium tissue were significantly increased (P<0.05), the expression levels of Bax and caspase-3 mRNA and proteins were significantly decreased(P<0.05), and the MDA levels and the activities of SOD, LDH and CK in serum had no significant differences(P>0.05). Compared with medium dose of crocin group, the expression levels of Bcl-2 mRNA and protein in myocardial tissue were significantly increased (P<0.05),the expression levels of Bax and caspase-3 mRNA and proteins were significantly decreased(P<0.05), the AI of myocardium of the rats in high dose of crocin group was significantly decreased (P<0.05), and the MDA level and the activities of SOD,LDH and CK in serum had no significant differences(P>0.05).

Conclusion

Different doses of crocin have protective effects on myocardial injury caused by ischemia and hypoxia in a dose-dependent manner, and its mechanism may be related to reducing the apoptosis of myocardial cells, and relieving myocardial oxidative stress injury.

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Inhibitory effects of siRNA targeting silencing TAK1 gene on proliferation and migration of thyroid cancer cells and p38 MAPK signaling pathway
Chunying ZHANG,Guangwei YIN,Mingda YOU,Hong CHEN,Yaojie HU,Yanbing LI,Chunyou CHEN
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  110-117.  DOI: 10.13481/j.1671-587x.20210115
Abstract ( 721 )   HTML ( 0 )   PDF (1079KB) ( 57 )  
Objective

To investigate the effects of siRNA targeting silencing transforming growth factor β-activated kinase 1 (TAK1) gene on the proliferation,migration and p38 mitogen-activated protein kinase (MAPK) signaling pathway of thyroid cancer cells,and to clarify the possible mechanism of silencing TAK1 gene in thyroid cancer.

Methods

Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting method were used to determine the expression levels of TAK1 mRNA and proteins in the thyroid cancer 8505C, NPA, BCPAP and KMH-2 cells. The KMH-2 cells were randomly divided into blank control group, negative control group and siRNA-TAK1 group. The cells in blank control group were not transfected,the cells in negative control group were transfected with negative control siRNA,and the cells in siRNA-TAK1 group were transfected with TAK1 siRNA. MTT method was used to measure the cell proliferation activity. Transwell assay was used to measure the invasion and migration abilities of cells. Western blotting method was used to determine the expression levels of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9(MMP-9),p38 and phosphorylated p38 (p-p38) proteins in the cells in various groups.

Results

Compared with the normal thyroid epithelial cells Nthyori3-1, the expression levels of TAK1 mRNA and protein in thyroid cancer 8505C, NPA, BCPAP and KMH-2 cells were significantly increased(P<0.05). Compared with blank control group and negative control group, the expression levels of TAK1 mRNA and protein in the KMH-2 cells in siRNA-TAK1 group were reduced (P<0.05), the cell proliferation activities were reduced (P<0.05), the number of invasion cells and migration cells was reduced (P<0.05),and the expression levels of PCNA, MMP-2, MMP-9 and p-p38 proteins in the cells were decreased (P<0.05). There were no significant differences in the indexes mentioned above of the KMH-2 cells between blank control group and negative control group(P>0.05).

Conclusion

SiRNA targeting silencing TAK1 gene can inhibit the proliferation,invasion and migration of thyroid cancer cells through inhibiting the activation of p38 MAPK signaling pathway.

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Improvement effect of tanshinone ⅡA on endometrial receptivity of intrauterine adhesion model rats and its mechanism
Jingqiao LIU,Yali MENG,Shuwen XU,Yuncan WANG,Na WANG,Yujing WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  118-124.  DOI: 10.13481/j.1671-587x.20210116
Abstract ( 652 )   HTML ( 0 )   PDF (1422KB) ( 34 )  
Objective

To observe the expression levels of integrin αVβ3 and leukemia inhibitory factor(LIF) in endometrium of the intrauterine adhesion(IUA) model rats, and to explore the effect of tanshinone ⅡA on endometrial receptivity of the IUA rats.

Methods

Fifty female SD rats were randomly divided into control group, model group, low dose of tanshinone ⅡA group, medium dose of tanshinone ⅡA group and high dose of tanshinone ⅡA group. The IUA rat models were established by curettage method. Seven days after molding, the rats in low, medium and high doses of tanshinone ⅡA groups were respectively given 5, 10 and 20 mg·kg-1 tanshinone ⅡA, while the rats in control group and model group were given equal volume saline once a day for 14 d. HE staining was used to observe the pathomorphology of endometrium tissue of the rats in various groups. Masson staining was used to detect the fibrotsis area ratios of endometrium of the rats in various groups. Immunohistochemistry method was used to observe the expression levels of integrin αVβ3 and LIF proteins in the endometrium tissue of the rats in various groups. Real-time quantitative PCR(RT-qPCR) was used to detect the expression levels of integrin αVβ3 and LIF mRNA in the endometrium tissue of the rats in various groups.

Results

The HE staining results showed the complete structure of uterine cavity, thick endometrium, abundant glands and orderly cell arrangement of the rats in control group; compared with control group, the cells in endometrium tissue of of the rats in model group were disorganized with fibrotic hyperplasia;compared with model group, the structures of uterine cavities of the rats in low, medium and high does of tanshinone ⅡA groups were obviously improved, the endometrium was thickened, the cell arrangement was in better order, and the fiber cells were decreased. Compared with control group, the number of endometrial glands of the rats in model group was significantly decreased (P<0.05), while the fibrosis area ratio was significantly increased (P<0.05), and the expression levels of integrin αVβ3 and LIF protein and mRNA were significantly decreased (P<0.05). Compared with model group, the number of endometrial glands in low, medium and high doses of tanshinone ⅡA groups were significantly increased (P<0.05), while the fibrosis area ratios were significantly decreased (P<0.05), and the expression levels of integrin αVβ3 and LIF protein and mRNA were significantly increased (P<0.05). Compared with low dose of tanshinone ⅡA group, the number of endometrial glands of the rats in medium and high doses of tanshinone ⅡA groups was significantly increased (P<0.05), while the fibrosis area ratios were significantly decreased (P<0.05), and the expression levels of integrin αVβ3 and LIF proteins and mRNA were significantly increased (P<0.05);compared with medium dose of tanshinone ⅡA group, the number of endometrial glands of the rats in high dose of tanshinone ⅡA group was significantly increased (P<0.05), while the fibrosis area ratios was significantly decreased (P<0.05), and the expression levels of integrin αVβ3 and LIF protein and mRNA were significantly increased (P<0.05).

Conclusion

Tanshinone ⅡA can improve the endometrial receptivity of the IUA rats by upregulating the expressions of integrin αVβ3 and LIF in endometrium tissue of the rats in a dose-dependent manner.

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Induction effect of gallic acid on apoptosis of human hepatocellular carcinoma HepG-2 cells through regulating mitochondrial oxidative phosphorylation
Haiying ZHANG,Wenxin ZHANG,Wenjing ZHAO,Wei LI
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  125-132.  DOI: 10.13481/j.1671-587x.20210117
Abstract ( 417 )   HTML ( 0 )   PDF (1380KB) ( 65 )  
Objective

To observe the inhibitory effect of gallic acid (GA) on the growth of human hepatocellular carcinoma HepG-2 cells,and to elucidate its mechanism.

Method

The human hepatocellular carcinoma HepG-2 cells were selected and treated with different concentrations (0, 3.125, 6.250, 12.500, 25.000, 50.000, and 100.000 mg·L-1) of GA and used as control group and 3.125, 6.250, 12.500, 25.000, 50.000,and 100.000 mg·L-1 GA groups, respectively. The survival rates of cells in various groups were detected by MTT assay. The HepG2 cells were divided into control group and 6.25, 12.50,and 25.00 mg·L-1 GA groups. The mitochondrial oxidative phosphorylation(OXPHOS)of Hep-G2 cells was analyzed using energy metabolism analyzer. Flow cytometry was used to detect the mitochondrial membrane potential (MMP) of the cells in various groups. Colorimetry was used to detect the adenosine triphosphate (ATP) levels in the cells in various groups. The expression amounts of cytochrome C (Cyt C) protein in cytoplasm and mitochondria in various groups were detected by Western blotting method. The morphology of cells was measured by acridine orange staining,and Annexin Ⅴ-FITC/PI double staining flow cytometry was used to detect the apoptotic rates of the cells in various groups.

Results

Compared with control group, the survival rates of HepG2 cells in different concentrations of GA groups were significantly decreased(P<0.05). Compared with control group, the basal respiration, maximum respiration capacities, respiration reserve capacities and ATP levels in 6.25, 12.50, and 25.00 mg·L-1 GA groups were significantly decreased (P<0.05);the MMP and ATP levels in the cells were significantly reduced (P<0.05); the expression amounts of Cyt C protein in the cytoplasm were increased, and the expression amounts of Cyt C protein in mitochondria were decreased; the cells in GA groups were shrank,the nuclei were shattered, and chromatin was aggregated; the early apoptotic rates and late apoptotic rates were increased significantly(P<0.05).

Conclusion

GA can inhibit the proliferation and induce the apoptosis of Hep-G2 cells,and its mechanism may be related to reducing MMP and inhibiting mitochondria OXPHOS.

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Effect of gestational diabetes mellitus on diaphragmatic function of newborn offsprings of rats
Ruili ZHANG,Xiaoqing YANG,Xiaoge ZHANG,Huafeng GUO,Jihong ZHU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  133-138.  DOI: 10.13481/j.1671-587x.20210118
Abstract ( 363 )   HTML ( 1 )   PDF (1318KB) ( 42 )  
Objective

To investigate the effects of gestational diabetes mellitus (GDM) on the diaphragmatic function and the oxidative stress levels of the newborn offsprings of the SD rats, and to clarify the possible mechanisms,

Methods

Forty-four pregnant female SD rats(aged 8 weeks) were randomly divided into normal control group and GDM model group. On the first day after pregnancy, the rats in GDM group were intraperitoneally injected with 35 mg·kg-1 streptozotocin(STZ) to establish the GDM rat models. The rats in normal control group were intraperitoneally injected with the equal volume of citric acid-sodium citrate buffer. After delivery, the diaphragm contractile force of the newborn rats in two groups was measured in vitro. Meanwhile,the cross section area of the diaphragm fiber was detected by immunofluorescence method. Routine pathological staining and transmission electron microscope (TEM) were used to observe the phathomorphology and ultrastructure of diaphragm tissue of the newborn offsprings of the rats. The levels of malondialdehyde(MDA) and carbonylated protein and the activities of superoxidase dismutase(SOD) and catalase(CAT) in the diaphragm tissue were measured with commercial kits.

Results

The results of contractile force detection in vitro showed that compared with normal control group, the diaphragmatic maximum tetanic force, maximum tension force, and contractile force under different stimulation frequencies of the newborn offsprings in GDM group were all significantly decreased(P<0.05), and the diaphragmatic fatigue index was increased (P<0.05). The immunofluorescence detection results showed that the cross section areas of type Ⅰ and type Ⅱ fibers of the newborn offspring rats in GDM group were significantly smaller than those in normal control group (P<0.05). The HE staining results showed the neatly and densely arranged diaphragm fibers, intact nuclear without obvious cellular swelling, and clear nucleus staining in diaphragm tissue of the newborn offspvings in two groups. The TEM observation results showed that the diaphragm fibers in normal control group were neatly arranged with regular and visible light-dark bands or “Z” lines, and the mitochondrial structure was intact with partitioned and visible ridges; however, the diaphragm fibers in GDM group were arranged with irregularly or even the light-dark bands and “Z” lines were disappeared,the amount of mitochondria was decreased,and the signs of mitochondrial apoptosis or vacuolation were found in diaphragm tissue of the newborn offsprings GDM group. Compared with normal control group, the levels of MDA and carbonylated protein in diaphragm tissue of the newborn offspring rats in GDM group were significantly increased(P<0.05), and the activities of SOD and CAT in GDM group were significantly decreased(P<0.05).

Conclusion

GDM can cause the decrease of diaphragmatic contractile force, reduction of cross section area of diaphragmatic fibers and changes of diaphragmatic ultrastructures,and the increase in oxidative stress level is one of the reasons that affect its diaphragm function.

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Effects of hirudin combined with atorvastatin on vascular endothelium function in rats with acute myocardial infarction
Ying CHENG,Jing YANG,Li RAO,Qiang XUE,Jie HU,Le FU,Qiaoyue ZHANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  139-144.  DOI: 10.13481/j.1671-587x.20210119
Abstract ( 992 )   HTML ( 4 )   PDF (975KB) ( 104 )  
Objective

To investigate the effects of hirudin combined with atorvastatin on the vascular endothelial function in the rats with acute myocardial infarction, and to elucidate the possible mechanisms.

Methods

A total of 50 rats were randomly divided into sham operation group, model group, hirudin group, atorvastatin group and combination group(n=10). The rat models with acute myocardial infarction were established by ligating the left anterior descending coronary artery.The rats in hirudin group, atorvastatin group and combination group were given 50 U·kg-1 hirudin, 10 mg·kg-1 atorvastatin, 50 U·kg-1 hirudin and 10 mg·kg-1 atorvastatin, respectively; while the rats in sham operation group and model group were given normal saline for 4 weeks. The 2,3,5-Triphenyte-trazoliumchloride (TTC) staining method was used to determine the myocardial infarction areas of the rats,and the percentage of myocardial infarction area was calculated. HE staining was used to observe the pathomorphology of myocardium tissue of the rats. Western blotting method was used to detect the protein expression levels of vascular cell adhesion molecule-1(VCAM-1) and intercellular adhesion molecule-1(ICAM-1) proteins in myocardium tissue of the rats. ELISA was used to determine the levels of endothelin-1(ET-1), von Willebrand factor (vWF), nitric oxide(NO), tumor necrosis factor (TNF-α )and interleukin- 6 (IL-6) in serum of the rats.

Results

Compared with sham operation group, the arrangement of myocardial cells in model group were disordered, some cells were incomplete, myocardial necrosis was serious, and inflammatory cell infiltration was found. Compared with model group,the arrangement of myocardial cells in hirudin group,atorvastatin group and combination group was in order,the myocardial necrosis was improved,and the inflammatory cell infiltraton was alleviated.Compared with sham operation group, the expression levels of VCAM-1 and ICAM-1 proteins in myocardium tissue of the rats in model group were significantly increased (P<0.05), the levels of ET-1, vWF, TNF-α and IL-6 in serum were significantly increased (P<0.05), and the level of NO was significantly decreased (P<0.05). Compared with model group, the percentages of myocardial infarction areas of the rats in hirudin group, atorvastatin group and combination group were significantly reduced (P<0.05), the expression levels of VCAM-1 and ICAM-1 proteins in myocardium tissue of the rats were significantly decreased (P<0.05),the levels of ET-1, vWF, TNF-α and IL-6 in serum were significantly decreased (P<0.05), and the levels of NO were significantly increased (P<0.05). Compared with hirudin group and atorvastatin group, the percentage of myocardial infarction area of the rats in combination group was significantly reduced (P<0.05), the expression levels of VCAM-1 and ICAM-1 proteins in myocardium tissue of the rats were significantly decreased (P<0.05),the levels of ET-1, vWF, TNF-α and IL-6 in serum were significantly decreased (P<0.05), and the level of NO was significantly increased (P<0.05).

Conclusion

Hirudin and atorvastatin can protect the vascular endothelium injury, and its mechanism may be related to regulating the secretion of active substances, inhibiting inflammation and reducing cell adhesion;combined therapy is more effective than hirudin or atorvastatin used alone.

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Inhibitory effect of curcumin on tumor growth in colorectal cancer mice and its mechanism of PTEN/PI3K/Akt signaling pathwayPEI Yongbin, WANG Guiqi, LI Wei, JIANG Xia, JIANG Haibo, ZHAO Zengren (Department of General Surgery,First Hospital,Hebei Medical University, Shijiazhuang 050031,China)
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  145-151.  DOI: 10.13481/j.1671-587x.20210120
Abstract ( 582 )   HTML ( 8 )   PDF (1409KB) ( 460 )  
Objective

To explore the effects of curcumin on the tumor growth and the phosphatase and tensin homologous genes deleted on chromosome ten/ phosphatidylinositol 3 kinase / protein kinase B (PTEN / PI3K / Akt) signaling pathway in the colorectal cancer mice,and to clarify the possible mechanism of curcumin in colorectal cancer.

Methods

A total of 60 mice were divided into model group, low dose of curcumin group, middle dose of curcumin group and high dose of curcumin group, with 15 mice in each group. The mice were inoculated with colorectal cancer LOVO cells to establish the mouse colorectal cancer models.The rats in low,middle and high doses of curcumin groups were given 25, 50 and 100 mg·kg-1 curcumin by gavage, respectively, for 4 weeks.Twenty four hours after the last administration,the tumor volumes, tumor weights and the inhibitory rates of tumor of the mice in various groups were detected. Immunohistochemical staining was used to detect the expressions of proliferating cell nuclear antigen (PCNA) in tumor tissue of the mice in various groups,and the proliferation index(PI) of cells was calculated. HE staining was used to observe the pathomorphology of tumor tissue of the mice in various groups, and Western blotting method was used to determine the expression levels of proto-oncogene c-Myc, caspase3, PTEN, Akt and phosphorylated Akt (p-Akt) proteins in tumor tissue of the mice in various groups.

Results

The tumor volumes, tumor weights, the inhibitory rates of tumor, PI values,and the expression levels of c-Myc, caspase3, PTEN and p-Akt proteins in tumor tissue of the mice in various groups had statistically significant differences (P<0.05).Compared with model group, the tumor volumes and tumor weights of the mice in different doses of curcumin groups were significantly decreased(P<0.05),the inhibitory rates of tumor were increased(P<0.05), the PI values were decreased(P<0.05),the expression levels of c-Myc and p-Akt proteins were decreased(P<0.05),and the expression levels of caspase3 and PTEN proteins were increased(P<0.05). The immunohistochemical results showed that the number of PCNA positive cells in tumor tissue of the mice in different doses of curcumin groups were significantly less than that in model group.The HE staining results showed that the tumor tissue cells in model group were disordered, the nucleus-cytoplasm ratio was increased, and the nuclear staining was deep; compared with model group, the tumor tissue cells of the mice in different doses of curcumin groups were relatively neatly arranged, the nucleus-cytoplasm ratios were decreased, and the nuclear staining was light.The effects of curcumin on tumor volume, tumor weight,inhibiory rate of tumor, PI value of tumor tissue and the expressions of c-Myc,caspase3,PTEN and p-Akt proteins in tumor tissue were dose-dependent, and there were statistically significant differences in the indicators among different doses of curcumin groups (P<0.05).

Conclusion

Curcumin can inhibit the tumor growth in the colorectal cancer mice,and its mechanism may be related to that curcumin can inhibit PTEN/PI3K/Akt signaling pathway.

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Inhibitory effect of sodium butyrate combined with X- irradiation on proliferation of lung cancer A549 cells and its influence in cell cycle
Guanhu LI,Qingxu LANG,Chunyan LIU,Qin LIU,Mengrou GENG,Xiaoqian LI,Zhenqi WANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  152-157.  DOI: 10.13481/j.1671-587x.20210121
Abstract ( 336 )   HTML ( 0 )   PDF (644KB) ( 72 )  
Objective

To study the inhibitory effect and radiosensitization effects of sodium butyrate (NaBt), a histone deacetylase inhibitor (HDACi), on the proliferation of human non-small cell lung cancer A549 cells alone or in combination with X-ray irradiation, and to analyze its combined effect with irradiation on the cell cycle of A549 cells.

Methods

The A549 cells in logarithmic growth phase were divided into control group, different concentrations (5, 10, 15, 20,and 25 mmol·L-1) of NaBt groups, 4 Gy X-ray irradiation group and different concentrations (5, 10, 15, 20,and 25 mmol·L-1) NaBt combined with 4 Gy X-ray irradiation groups. CCK-8 method was used to detect the cell proliferation rates in various groups at 24, 48 and 72 h after treatment. The changes of cell cycle were analyzed by flow cytometry.The A549 cells in logarithmic growth phase were divided into control group, 10 mmol·L-1 NaBt group,20 mmol·L-1 NaBt group, 4 Gy X-ray irradiation group, 10 mmol·L-1 NaBt combined with 4 Gy X-ray irradiation group, and 20 mmol ·L-1 NaBt combined with 4 Gy X-ray irradiation group. At 24 h after treatment, the percentages of A549 cells in each phase of cell cycle in various groups were analyzed.

Results

After 48 h of treatment, the cell proliferation rates in 5,10, 15, 20, and 25 mmol·L-1 NaBt groups were significantly lower than that in control group(P<0.05 or P<0.01);after 72 h of treatment, the cell proliferation rates in 5,10,20,and 25 mmol·L-1 NaBt groups were significantly lower than that in control group(P<0.01) in a concentration- and time-dependent manner;after treated for 48 h, the cell proliferation rates in 15, 20,and 25 mmol·L-1 NaBt combined with 4 Gy X-ray irradiation groups were significantly lower than those in control group,4 Gy X-ray irradiation group and corresponding concentrations of NaBt groups (P<0.05 or P<0.01);after treated for 72 h,compared with control group and 4 Gy X-ray irradiation group,the cell proliferation rates in different concentrations of NaBt combined with 4 Gy X-ray irradiation groups were significantly decreased(P<0.01);the cell proliferation rates in 20 and 25 mmol·L-1 NaBt combined with 4 Gy X-ray irradiation groups were significantly lower than those in corresponding concentrations of NaBt groups(P<0.01).The cell cycle analysis results showed that after 24 h of treatment, compared with control group, the percentages of A549 cells in G0 / G1 phase and G2 + M phase in 10 and 20 mmol·L-1 NaBt groups were significantly increased (P<0.05 or P<0.01), and the percentage of A459 cells in S phase was significantly decreased (P<0.01); the percentages of cells in G2+M phase in 4 Gy X-ray irradiation group and 10 mmol·L-1 NaBt combined with 4 Gy X-ray irradiation group were significantly increased (P<0.05 or P<0.01),and the percentages of cells in G0/G1 phase and S phase were significantly decreased (P<0.01).

Conclusion

NaBt has the inhibitory effect on the proliferation of A549 cells and radiosensitization effect in a concentration- and time-dependent manner, and the combined inhibitory effect of NaBt and X-ray is better than both of them used alone; its mechanism may be related to G2 + M phase arrest of the A549 cells induced by combination of NaBt and X-ray.

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Inhibitory effect of baicalin on inflammation in rats with spinal cord injury by regulating macrophage M2 polarization
Dayong XU,Yunpeng LI,Jingmei WEI,Ruyin LIU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  158-167.  DOI: 10.13481/j.1671-587x.20210122
Abstract ( 700 )   HTML ( 4 )   PDF (1128KB) ( 172 )  
Objective

To explore the protective effect of baicalin on the spinal cord injury (SCI) in the rats, and to clarify its mechanism.

Methods

The SCI models of rats were established by modified Allen’s method in vivo. Fifty healthy SD rats were randomly divided into sham operation group (given normal saline), model group (modeling, given normal saline), baicalin group (modeling, given 200 mg·kg-1 baicalin), baicalin+ DMSO group (modeling, given 200 mg·kg-1 baicalin and 0.2% DMSO) and baicalin +AS1517499 (STAT6 pathway inhibitor) group (modeling, given 200 mg·kg-1 baicalin and 10 mg·kg-1 AS1517499)(n=10). After the models were successfully established, the spinal cord motor function of rats was evaluated by using the Basso-Beattie-Bresnahan (BBB) method. In vitro, 2.5 μg·L-1 lipopolysaccharide (LPS) and 0.125 μg·L-1 interferon-γ (IFN-γ) were used to induce M1 polarization of macrophages (RAW264.7), and 10 μg·L-1 interleukin-4 (IL-4) was used to induce M2 polarization of macrophages. The RAW264.7 cells were divided into control group, M1 type polarization group, M2 type polarization group,M1 type polarization+ baicalin group, M2 type polarization+ baicalin group, M1 type polarization+baicalin+AS1517499 group,and M1 type polarization+baicalin+DMSO group. ELISA assay was used to detect the levels of inflammatory factors interleukin-12(IL-12),tumor necrosis factor-α(TNF-α),interleukin-4( IL-4) and interleukin-10(IL-10) in the spinal cord tissue and RAW264.7 cells of the rats; Western blotting method was used to detect the expression levels of M1 type macrophage marker proteins inducible nitric oxide synthase(iNOS)and chemokine-5( CCL-5), M2 type macrophage marker proteins arginase 1(Arg1)and mannose receptor C1(MRC1) and Janus kinase 1/signal transducer and activator of transcription 6(JAK1/STAT6) pathway related proteins in spinal cord tissue and RAW264.7 cells of the rats.

Results

In vivo experiments, compared with model group, the BBB score of the rats in baicalin group was significantly increased (P<0.05), the levels of IL-12 and TNF-α and the expression levels of iNOS,CCL-5,and p-STAT1 proteins in spinal cord tissue were significantly reduced (P<0.05), and the levels of IL-4 and IL-10 and the expression levels of Arg1, MRC1, p-JAK1 and p-STAT6 proteins were significantly increased (P<0.05). Compared with baicalin+DMSO group, the levels of IL-12 and TNF-α and the expression levels of iNOS, CCL-5 and p-STAT1 proteins in the spinal cord tissue of the rats in baicalin+AS1517499 group were significantly increased (P<0.01), and the levels of IL-4 and IL-10 and the expression levels of Arg1, MRC1,and p-STAT6 proteins were significantly reduced (P<0.01). In vitro experiments, compared with M1 type polarization group, the expression levels of iNOS, CCL-5 and p-STAT1 proteins in the RAW264.7 cells in M1 type polarization + baicalin group were significantly reduced (P<0.05),and the expression levels of p-JAK1 and p-STAT6 proeins were significantly increased(P<0.05). Compared with M2 type polarization group, the expression levels of Arg1 and MRC1 in M2 type polarization+baicalin group were significantly increased (P<0.05). Compared with M1 type polarization+baicalin+DMSO group, the levels of IL-12 and TNF-α in the RAW264.7 cells in M1 type polarization+baicalin+AS1517499 group were significantly increased (P<0.05), and the levels of IL-12 and TNF-α were significantly decreased (P<0.05), the expression levels of p-STAT6, Arg1 and MRC1 proteins were significantly reduced (P<0.05), and the expression levels of iNOS and CCL-5 proteins were significantly increased(P<0.05).

Conclusion

Baicalin could activate the JAK1/STAT6 pathway and inhibit the inflammatory process of SCI rats by promoting M2 polarization of macrophages.

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Research in clinical medicine
Changes in levels of brain injury markers in serum during perioperative period in infants underwent parent liver transplantation and their clinical significances
Hongli YU,Wenli YU,Yiqi WENG,Weihua LIU,Ying SUN,Yunxia LIU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  168-172.  DOI: 10.13481/j.1671-587x.20210123
Abstract ( 315 )   HTML ( 0 )   PDF (650KB) ( 158 )  
Objective

To investigate the changes in the levels of serum neuron specific enolase (NSE) and serum S-100β protein during the perioperative period in the infants underwent parent liver transplantation, and to clarify the relationships between NSE, S-100β and brain injury during the perioperative period of the infants underwent parent liver transplantation.

Methods

Forty infants with congenital biliary atresia underwent parent liver transplantation were selected, aged 4-12 months, weighing 3.0-10.0 kg, ASA status Ⅲ or Ⅳ class. The blood samples were drawn from central vein of the infants before skin incision (T1), 30 min after anhepatic phase (T2), 1 h after neohepatic phase (T3) and 24 h after neohepatic stage (T4). Furthermore, the heart rate (HR), the mean arterial blood pressure(MAP), the central venous pressure(CVP),and pH values of blood were monitored at the moments of T1-T 4. The serum S-100β and NSE levels were detected by ELISA method.

Results

The levels of hemodynamics were significantly changed in anhepatic phase and neohepatic phase;compared with T1,the HR at T2 of the infants was accelerated (P<0.05),and the MAP, CVP and pH value of blood were decreased (P<0.05). After the surgery, all the hemodynamic indexes fell to the preoperative levels. Compared with T1, the serum NSE and S-100β levels at T2-T 4 of the infants were decreased (P<0.05); compared with T2, the serum S-100β and NSE levels of the infants at T3 of the infants were significantly increased (P<0.05); compared with T3,the serum S-100β and NSE levels at T4 of the infants were significantly decreased (P<0.05).

Conclusion

The brain injury may appear during the perioperative in the infants underwent parent liver transplantation and gradually aggravates during the anhepatic phase and gradually returns to the preoperative levels at 24 h after surgery.

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Effects of EDC on immediate and aging micro-tensile bonding strength of dentin
Qing SHA,He LI,Hong ZHANG,Boqun CHENG,Ying WANG,Linlin YAN,Xinying ZOU,Zhimin ZHANG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  173-179.  DOI: 10.13481/j.1671-587x.20210124
Abstract ( 359 )   HTML ( 0 )   PDF (736KB) ( 86 )  
Objective

To explore the the cytotoxicity, micro-tensile bond strength, and fracture mode of the universal adhesive after added with 1-ethyl-3 (3- dimethylaminopropyl) carbodiimide(EDC),and to clarify the effect of EDC addition amount on the bonding performance of universal adhesive.

Method

EDC was added to the universal adhesive Single Bond Universal to configure the experimental adhesives containing different concentrations (0.1,0.2 and 0.3 mol·L-1 ) of EDC.The L-929 fibroblasts in the logarithmic phase were divided into negative control group and 0,0.1,0.2 and 0.3 mol·L-1 EDC groups, and blank control group (culture medium) was set up;the L-929 cells in negative control were added with culture solution,and the adhesive film was prepared and the extract was obtained in different concentrations of EDC groups .The extract was co-cultured with the cells.The relative growth rate (RGR) was measured by CCK-8 method, and the cytotoxicity was graded.The freshly extracted third molars without caries were selected to prepare the micro-tensile bonding specimens. The universal testing machine was used to test the immediate(the specimens were stored in 37 ℃ deionized water for 24 h ) and aging(the specimens were treated in the hot and cold cycle machine for 5 000 times of circulation) micro-tensile bonding strengths of the specimens in various groups.The immediate micro-tensile test included control group (without EDC) and 0.1, 0.2 and 0.3 mol·L-1 EDC groups,and the aging micro-tensile test included control group (without EDC), and 0.1 and 0.2 mol·L-1 EDC groups.The fracture mode of the specimens was observed with a stereo microscope.

Results

In the cytotoxicity experiment, the RGR of cells in various group at 24, 48 and 72 h after co-cultivation were all above 75%,and the cytotoxicity was level 1. In the immediate micro-tensile test, the bonding strengths in control group and 0.1, 0.2 and 0.3 mol·L-1 EDC groups were (30.71±4.36), (39.41±6.72), (26.51±9.54) and(18.55±5.37) MPa.Compared with contol group,the bonding strength in 0.1 mol·L-1 EDC group was significantly increased (P<0.05),the bonding strength in 0.3 mol·L-1 EDC group was significantly reduced (P<0.05). The fracture mode observation results showed that the interface failure was the main performance in control group, 0.2 and 0.3 mol·L-1 EDC groups,and the mixed failure was the main performance in 0.1 mol·L-1 EDC group. In micro-tensile test,the bonding strengths in control group,0.1 mol·L-1 EDC group and 0.2 mol·L-1 EDC group were (21.42±1.58), (35.70±1.17), and (16.24±3.27) MPa, respectively, Compared with control group, the bonding strength in 0.1 mol·L-1 EDC group was significantly increased (P<0.05),and the bonding strength in 0.2 mol·L-1 EDC group was significantly reduced (P<0.05). The fracture mode observation results showed that the interface failure was the main performance in control group and 0.2 mol·L-1 EDC group, and the mixed failure was the main performance in 0.1 mol·L-1 EDC group.

Conclusion

The experimental adhesive containing 0.1 mol·L-1 EDC after added with EDC to the Single Bond Universal has no obvious cytotoxicity, and the immediate and aging micro-tensile bonding strengths are improved.

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Genomic evolution and drug resistance analysis of human-derived Listeria monocytogenes
Guang YAO,Yuying LU,Qinghua ZHANG,Haixia ZHU,Yiwei CHEN,Dong SUN,Zhen ZHUANG,Feng ZHANG,Ding LIU,Zhi SONG
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  180-186.  DOI: 10.13481/j.1671-587x.20210125
Abstract ( 421 )   HTML ( 0 )   PDF (1649KB) ( 99 )  
Objective

To identify the Listeria monocytogenes(LM) obtained from the cerebrospinal fluid of one patient with meningitis with new generation sequencing technology and analyze genomic evolution and drug resistance, and to explore the possible causes of rapid progression of disease and treatment failure.

Methods

The LM were cultivated, isolated and identified from the patient’s cerebrospinal fluid, and bacterial De novo sequencing was conducted by the Illumina HiSeq 3000 system to obtain the whole genome data of the strain.Furthermore, the evolutionary tree of the LM was constructed and the drug-resistant related genes were identified by gene assembly, gene prediction, gene annotation, single nucleotide polymorphism(SNP) analysis, and phylogenetic analysis and other bioinformatics methods.

Results

The bacteria in the cerebrospinal fluid were isolated and cultured, and identified as LM by mass spectrometry.The size of the assembled bacterial genome after sequencing was 3 008 507 bp.By aligned with database, its genome was accordance with 88% of LM genome, confirming that the bacteria was LM.The obtained listeria strains were close to FSL N1-017 and SLCC2540 by aligning 38 representative listeria genome construction phylogenetic trees in the NCBI database,and they had no evolutionary relationships with human-derived LM that were reported worldwide. It was considered to be a newly discovered clinically infectious LM in China.The drug resistance analysis revealed that the obtained LM genome contained some drug-resistant genes associated with the major facilitator superfamily(MFS)transporters,and four of them were matched the drug resistance genes in the database EDG-e strain transporter annotation, suggesting that the strain had strong clinical resistance.

Conclusion

The cultured bacteria from cerebrospinal fluid of clinical meningitis patient is identified as a new type of clinically infectious pathogenic LM strain with strong clinical drug resistance.

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Expressions of miRNA-508-3p and HGF in placenta tissue of patients with gestational diabetes mellitus and their effects on trophoblast insulin resistance
Hao HUANG,Hong JIA,Xiaoshuang WANG,Lu ZHANG,Yating DUAN
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  187-195.  DOI: 10.13481/j.1671-587x.20210126
Abstract ( 423 )   HTML ( 0 )   PDF (1277KB) ( 67 )  
Objective

To investigate the expressions of microRNA-508-3p (miR-508-3p) and hepatocyte growth factor (HGF) in placenta tissue of the patients with gestational diabetes mellitus (GDM) and their relationship, and to clarify the related molecular mechanism of miR-508-3p in mediating trophoblast insulin resistance(IR).

Methods

A total of 15 GDM patients and 15 normal pregnant women were selected and used as GDM group and normal control group. The expression levels of miR-508-3p and HGF protein in placenta tissue of the patients in two groups were detected by RT-PCR, immunohistochemistry and Western blotting methods, and their relationship was analyzed.HGF was screened as the target gene of miR-508-3p with bioinformatics,which was verified by double luciferase reporter gene assay and Western blotting method. The human trophoblast cells HTR-8/Svneo were cultured in vitro to establish the HTR-8/Svneo IR cell model (HTR-8/Svneo-IR). The HTR-8/Svneo-IR cells were transfected with miR-508-3p-inhibitor (miR-508-3p-inhibitor-IR grouop) and miR-NC(miR-NC-IR group) by liposome transient transfection method. The transfection efficiency was detected by RT-PCR; Western blotting method was used to detect the expression levels of HGF and phosphorylated phosphatidylinosital 3 kinase(p-PI3K) and phosphorylated protein kinse B(p-AKT) proteins in PI3K/AKT signaling pathway in the cells in various groups.

Results

Compared with normal control group, the expression levels of miR-508-3p in placenta tissue of the patients in GDM group was significantly increased (P<0.05), while the HGF protein expression level was significantly decreased (P<0.01), and there was a negative correlation between the expression level of HGF protein and the expression level of miR-508-3p in placenta tissue of the patients in GDM group(r =-0.542, P<0.05). Compared with HTR-8/Svneo group, the expression level of miR-508-3p in the cells in HTR-8/Svneo-IR group was significantly increased (P<0.05);compared with HTR-8/Svneo-IR group,the expression level of miR-508-3p in the cells in miR-508-3p-inhibitor-IR group was significantly decreased (P<0.05).Compared with HTR-8/Svneo group, the expression levels of HGF,p-PI3K and p-AKT proteins in the cells in HTR-8/Svneo-IR group were significantly decreased (P<0.05);compared with HTR-8/Svneo-IR group, the expression levels of HGF,p-PI3K and p-AKT proteins in the cells in miR-508-3p-inhibitor-IR group were significantly increased (P<0.05).

Conclusion

MiR-508-3p can inhibit the expression of HGF in placenta tissue of the GDM patients, and promote IR of trophoblasts through PI3K/AKT signaling pathway, and then participate in the occurrence and development of GDM.

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Clinical medicine
Pulmonary epithelioid hemangioendothelioma complicated with chronic obstructive pulmonary disease:A case report and literature review
Jing HUANG,Ming DING,Xiaoli ZHU,Pingsheng CHEN,Shuhua HAN
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  196-202.  DOI: 10.13481/j.1671-587x.20210127
Abstract ( 559 )   HTML ( 7 )   PDF (1190KB) ( 81 )  
Objective

To analyze the clinical manifestation, diagnosis and treatment of the patient with pulmonary epithelioid hemangioendothelioma (PEHE) combined with chronic obstructive pulmonary disease(COPD), and to develop the clinicians’ understanding of PEHE.

Methods

The clinical data of a patient with PEHE complicated with COPD were collected; the relevant literatures were reviewed, and its diagnosis and treatment methods were summarized.

Results

The male patient was 74 years old, who was hospitalized because of repeated chest tightness and asthma for half a year and half a month of deterioration. Physical examination showed barrel chest, and hyperresonant sounds were heard when percussing lungs; the respiratory sounds of lungs were reduced; there were no other obvious positive signs. The examination results of pulmonary function showed that forced expiratory volume in 1 second (FEV1) accounted for 58% of predicted value and FEV1/ forced vital capacity (FVC) was 56.9%. The chest CT results showed multiple nodules with different sizes, chronic bronchitis and bullous emphysema in both lungs. The bronchoscope results showed abnormal cell-nest, the clinical and immunologic markers were considered, and the results met the diagnosis of PEHE. The patient got better after treated with endostar and bronchodilator and was discharged from hospital. After that, chest tightness and asthma occurred again, so the patient was admitted to the hospital for symptomatic treatment because of COPD, but the symptoms were not relieved significantly and fatigue and systemic pain were found after endostar was given. The evaluation of imaging revealed intrahepatic metastases,and bone metastases in several areas, so the patient received anti-tumor and symptomatic treatment by taking anlotinib. Then, respiratory failure, hypoalbuminemia and secondary infection occurred. So symptomatic treatment was given after assessing the lesions of both lungs by imaging. But the patient was dead.

Conclusion

The clinical symptoms of PEHE are atypical, and it is easy to be ignored when a patient is accompanied by COPD. The presence of COPD can also affect what kind of treatment plan can be used.

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Imageology
Effects of three-dimensional transrectal ultrasound and MRI in evaluation of extramural vascular invasion degrees of middle and lower rectal cancer
Dong CHEN,Haitao CHEN,Zhiyao LI,Fengming RANG,Xi ZHANG,Zhirui CHUAN,Shicong TANG,Xiaomao LUO
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  203-209.  DOI: 10.13481/j.1671-587x.20210128
Abstract ( 679 )   HTML ( 0 )   PDF (841KB) ( 44 )  
Objective

To compare the results of three-dimensional endorectal ultrasound (3D-ERUS) and magnetic resonance imaging (MRI) in the diagnosis of extramural vascular invasion (EMVI) of middle and lower rectal cancer before operation,and to explore the clinical value of 3D-ERUS in the evaluation of EMVI of middle and lower rectal cancer before operation.

Methods

A total of 94 patients with rectal cancer confirmed by pathology were selected,and all patients were examined by 3D-ERUS and MRI. The 3D-ERUS and MRI images were evaluated with a grade 5 scores (0-4 scores) for EMVI.Receiver operating characteristic(ROC) curve was used to evaluate the results of 3D-ERUS and MRI in the diagnosis of EMVI, and the specificity (Sp),sensitivity (Se),positive predictive value (PPV),negative predictive value (NPV) accuracy,and the area under ROC curve(AUC) were calculated. Kappa test was used to evaluate the consistency between the results of 3D-ERUS and MRI in the diagnosis of EMVI and the pathological results.

Results

The accuracy of 3D-ERUS in the diagnosis of EMVI was 90.43%(Kappa=0.685,P<0.01), AUC was 0.868, Se was 81.25%,Sp was 92.31%,PPV was 68.42% and NPV was 96.00%.The accuracy of MRI in the diagnosis of EMVI was 87.23%(Kappa=0.589,P<0.01),AUC was 0.824, Se was 75.00%,Sp was 89.74%,PPV was 60.00% and NPV was 94.59%. There was no statistical difference in the accuracy of diagnosis between 3D-ERUS and MRI(χ2=0.214,P=0.643),and there was no statistical difference in the AUC of diagnosis between 3D-ERUS and MRI(Z=1.355,P=0.175).

Conclusion

3D-ERUS and MRI eualuation on EMVI of the patients with middle and lower rectal cancer before operation have highly clinical values. 3D-REUS can be used as a complementary tool of MRI to evaluate the EMVI of the patients with middle and lower rectal cancer before operation.

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Survey research
Detection of DNA in spotted fever group Rickettsia carried by Dermacentor nuttalli in partial areas of Inner Mongolia and its distribution of genotypes
Zheng GUI,Jingfeng YU,Lan MU
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  210-215.  DOI: 10.13481/j.1671-587x.20210129
Abstract ( 633 )   HTML ( 1 )   PDF (991KB) ( 50 )  

Objective: To investigate the status of spotted fever group Rickettsia (SFGR) carried by Dermacentor nuttalli in partial areas of in Inner Mongolia, and to analyze the species of tick-carried SFGR in this area and perform the homology analysis.

Methods

In the middle of April 2019, 264 Dermacentor nuttallis from 708 sheep were collected from Chengchuan Town, early Banner of Etoke Banner, Erdos City; Siziwang Banner, Hohhot City and Bayan WenduSumu area, Arukorqin Banner, Chifeng City, Inner Mongolia. After DNA extraction of single tick, Rickettsia 16sRNA was amplified by PCR method as a preliminary screening experiment, ten positive samples were randomly selected from each region, and a total of 30 positive samples were further amplified for gltA and ompA genes, then the positive samples were sequenced and cluster analysis was performed.

Results

Among 264 ticks, 218 were positive SFGR, with the positive rate of 82.57%; 14 SFGR ompA positive samples and 7 SFGR gltA positive samples were sequenced successfully. The similarities of gltA gene and ompA gene were 100% and 99.86%. The results of phylogenetic analysis showed that the detected sequence was in the same branch with Rickettsia raoultii, and the gltA gene sequence was closely related to Candidatus Rickettsia uralicaRickettsia parkeri and Rickettsia sibirica, while the ompA gene sequence was close to that of Rickettsia massiliae and Rickettsia rhipicephali; both of them were far away from Rickettsia monacensis.

Conclusion

The genotype of SFGR carried by Dermacentor nuttalli in Ordos, Siziwang Banner and Chifeng, Inner Mongolia is Rickettsia raoultii.

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Mediating effect of perceived social support between self-esteem and anxiety in patients with coronary heart disease
Yang YU,Yang YANG,Mingtu XU,Zeying QIN,Cong FU,Jingyang LI
Journal of Jilin University(Medicine Edition). 2021, 47 (1):  216-221.  DOI: 10.13481/j.1671-587x.20210130
Abstract ( 335 )   HTML ( 2 )   PDF (692KB) ( 117 )  
Objective

To explore the anxiety and its related factors in the patients with coronary heart disease,and to investigate the mediating effect of perceived social support between self-esteem and anxiety.

Methods

A total of 315 patients with coronary heart disease who were first admitted to hospital were investigated by using the basic information questionnaire, General Anxiety Disorder-7, Self-Esteem Scale and Perceived Social Support Scale in the study. A total of 305 valid questionnaires were obtained. The influencing factors related to anxiety of the patients were compared between two groups and multiple groups with the method of t-test and single factor analysis of variance.Pearson correlation was used to analyze the correlations between self-esteem, perceived social support and anxiety. Stepwise regression analysis was used to test the mediating effect.

Results

The anxiety level of female in the patients with coronary heart disease was higher than that of male (t=9.664,P<0.01). The anxiety levels of the patients with different education levels were statistically significant (F=3.146,P<0.05), and the anxiety level of the illiterate patients was the highest. The anxiety levels of the patients with different marital status were statistically significant(F=9.113,P<0.01), and the anxiety level of widowed patients was the highest. The anxiety level of the patients with sleep disorders was higher than that of the patients without sleep disorders (t=19.961, P<0.01). The anxiety level of the patients with alcohol addiction was higher than that of the patients without alcohol addiction (t=10.462, P<0.01). Self-esteem was positively correlated with perceived social support (r=0.251, P<0.01), self-esteem was negatively correlated with anxiety (r=-0.173, P<0.01), and perceived social support was negatively correlated with anxiety (r=-0.187, P<0.01). Self-esteem could directly predict anxiety (β=-0.224, P<0.01), and perceived social support played a partial mediating role between self-esteem and anxiety(β=-0.224, P<0.01;β=-0.070, P<0.05).

Conclusion

Anxiety problems are prominent in the female, illiterate, widowed, alcohol addiction and sleep-disordered patients with coronary heart disease. Low self-esteem and poor perceived social support levels are the predictors of anxiety,and perceived social support plays a partial mediating role between self-esteem and anxiety.

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