Table of Content

05 March 2020, Volume 40 Issue 3

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Original Articles

Recombinant human erythropoietin improves neurological function after traumatic brain injury of mice

WU Chen-rui, HE Hong-bing, CHENG Yu-qi, MAO Lan-tian, LIAO Zheng-bu

2020, 40(3):  289-293. 

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Objective To investigate the effects of recombinant human erythropoietin (rhEPO) on angiogenesis and neurofunction recovery of mice with traumatic brain injury(TBI). Methods Mice were randomly divided into sham operation group (sham group), traumatic brain injury group (TBI group, severe TBI model made by controlled cortical impact) and rhEPO treatment group (EPO group, rhEPO intraperitoneal injected at 5 000 IU/(kg·d) for 7 days after the trauma). Each group included 15 mice. Neurological scores were performed at 3, 7, and 14 d using modified neurological severity score(mNSS). Fourteen days after injury, Western blot and immunofluorescence were used to detect the protein expression of eNOS, VEGF and CD31 around ipsilateral cerebral cortex, and the microvessel density (MVD) was detected by CD31⁺ cells. The lesion volume was measured at 21 d after injury using hematoxylin-eosin (HE) stained specimens. Results 3,7 and 14 d after TBI, mNSS of TBI group was signifi- cantly higher than that of sham group (P＜0.001). 7 and 14 d after TBI, mNSS of EPO group was lower than those of TBI group (P＜0.05). Fourteen days after TBI, the protein expression of eNOS, VEGF and CD31 was higher in TBI group than those of sham group, and the EPO group was higher than those of TBI group. (P＜0.05). 14 d after TBI, the MVD of TBI group was significantly lower than sham group(P＜0.001), and the MVD of EPO group was significantly higher than TBI group(P＜0.001). Twenty one days after injury, the volume of EPO group was reduced compared with TBI group (8.90±1.79) (P＜0.05). Conclusions rhEPO promotes angiogenesis and improves neurological function after TBI of mice.

Regulation of co-transcription factor CITED1 in mouse bone metabolism

QIU Yi-yan, ZHENG Yu-cheng, GUO Wei-zhuang, ZHOU Wen-yu, YAN Bin, YANG Xin-jian

2020, 40(3):  294-298. 

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Objective To study the regulation of CITED1 in the mice bone metabolism, including osteogenesis/osteoclast balance in vivo to provide a theoretical basis for the treatment of osteoporosis. Methods Bone phenotypes such as femur length, bone mass, bone cortex and bone cancellous thickness of KO mice were measured by micro CT. Hematological related parameters of bone metabolism were determined by ELISA. RT-qPCR was used to detect the expression of bone marker genes to explore the causes of bone metabolism changes after CITED1 gene knockout. Results The expression of CITED1 was low in KO mice, indicating a successful CITED1 knockout. Femoral length, bone mass, bone cortex and bone cancellous thickness were significantly higher in KO mice than those in WT mice. The concentration of type I procollagen peptide (P1NP), osteocalcin (OC) and bone alkaline phosphatase (BALP) in serum of KO mice was significantly higher than those of WT mice, while the concentration of tartaric acid phosphatase (TRAP) was significantly lower. The expression of OC and BALP in KO mice was significantly higher than that in WT mice(P＜0.001). At the same time the expression of TRAP in KO mice was signifi- cantly lower(P＜0.05). Conclusions CITED1 knockout can up-regulate the expression of OC and BALP, and down-regulate the expression of TRAP for promoting bone formation and inhibiting bone resorption.

Curcumin alleviates deep tissue pressure injury in mice

LIU Pan-pan, ZHANG Zi-rui, YANG Xu, HAN Jing, LU Lian-fang, WANG Ai-min, ZHANG Ju

2020, 40(3):  299-304. 

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Objective To evaluate the effect of curcumin on wound healing in deep tissue pressure injury(DTPI) induced by ischemia-reperfusion in mice. Methods Mice were randomly divided into control group (Con), model group (Mod), 4 and 8 mg/kg curcumin treatment group (Cur 4, Cur 8), 10 mice in each group. A deep tissue pressure injury model was established by magnet compression. The wounds tissue was injected with curcumin every 48 hours and photographed to observe the wound healing. HE staining was used to observe the pathological changes of wound healing. Real-time PCR was used to detect the mRNA expression of TNF-α, IL-6, IL-10, VEGF-α and TGF-β. Western blot was used to detect the expression of VEGF-α, TGF-β, Stat3 and p-Stat3. Results Compared with the control group, the wound in the model group gradually deteriorated, and the pathological changes were obvious. Compared with the model group, the wound healing of the Cur 8 group was significantly accelerated (P＜0.05). Compared with the model group, the inflammatory cell infiltration was reduced and the neovascular density was increased in the Cur 8 group. The mRNA expression of TNF-α and IL-6 in the wound tissues of the Cur 8 group was less than those in the model group, while the expression of IL-10, VEGF-α and TGF-β was higher than those in the model group (P＜0.05). The expression of VEGF-α, TGF-β and p-Stat3 in Cur 8 group was higher than those in model group (P＜0.05). Conclusions Curcumin administration can alleviate DTPI of mice induced by ischemia-reperfusion, and its mechanism may be related to the inhibition of inflammatory response and promotion of angiogenesis.

Estrogen promotes proliferation and invasion of astrocyte cell line U-87

LIU Yan-min, FEI Yi-lin, LI Xing-miao, WANG Ning, CHEN Mo, JIANG Xiu-xiu

2020, 40(3):  305-309. 

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Objective To explore the relationships between the estrogen and astrocyte cell proliferation as well as invasion. Methods Astrocyte cell line U-87 was cultured in vitro and treated with 0, 10^－12, 10^－10, 10^－8 and 10^－6 mol/L estrogen. The gene and protein expressions were analyzed by qPCR, Western blot, ELISA; and the cell proliferation and invasion was analyzed by CCK-8 kits and transwell experiments. Results Estrogen significantly inhibited IL-1, TNF-α, IL-6 expression and promoted the expression of NGF and PGE2 (P＜0.05). Furthermore, estrogen inhibited aquaporin 2(AQP2) expression (P＜0.05). The proliferation and invasion experiments showed that estrogen promoted the U-87 cell proliferation and invasion (P＜0.05). Conclusions Estrogen down-regulates the expression of inflammatory factors, inhibites the expression of AQP2, promotes the proliferation and invasion of U-87 cells.

Oxaliplatin combined with daunorubicin induces apoptosis of colon cancer cell line HT-29

TIAN Chun-yang, LIU Xiao-zheng, FAN Jun-chao

2020, 40(3):  310-314. 

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Objective To study the mechanism of oxaliplatin combined with daunorubicin on the apoptosis of colon cancer cells. Methods Colon cancer cells HT-29 were incubated with different concentrations of oxaliplatin and daunorubicin respectively, MTT assay was used to detect cell proliferation and calculate the median inhibitory concentration.HT-29 cells were treated with half inhibitory concentration of oxaliplatin, daunorubicin and by two drugs combined. MTT assay cell proliferation ability, the colony forming ability of cells was detected by plate cloning test, cell apoptosis was detected by flow cytometry, Western blot was used to detect the expression of cleaved caspase-3, PTEN and p-Akt in cells. Results Different concentrations of oxaliplatin and daunorubicin inhibited the proliferation of HT-29 cells, the half inhibitory concentration of daunorubicin was about 0.23 μmol/L and oxaliplatin was about 40 mg/L. Daunorubicin, oxaliplatin or oxaliplatin plus daunorubicin were used to treat HT-29 cells, the proliferation of cells decreased, the ability of clone formation also reduced, the rate of apoptosis increased, the expression of cleaved caspase-3 protein increased in cells, the expression of p-Akt protein in cells decreased,the expression of PTEN protein elevated(P＜0.05). The effect of oxaliplatin combined with daunorubicin was higher than that of daunorubicin or oxaliplatin alone (P＜0.05). Conclusions Oxaliplatin combined with daunorubicin can induce apoptosis in colon cancer cells, its mechanism may be related to the regulation of PTEN/Akt signaling.

Triptolide inhibits the proliferation and promotes apoptosis of non-small cell lung cancer cell line A549

LIU Qian, TANG Jian-hua, ZHANG Zhi-hua

2020, 40(3):  315-320. 

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Objective To investigate the effect of triptolide (TP) on the proliferation and apoptosis of human non-small cell lung cancer cell A549 by regulating the expression of chemokine receptor 4 (CXCR4) gene. Methods The experiment was divided into 4 groups: control group, TP group (100 nm/L TP-treated cells), CXCR4+TP group (transfected plasmid TP-treated cells) and NC+TP group (empty plasmid TP-treated cells). The expression of CXCR4 and transfection were detected by RT-qPCR and Western blot. Cell proliferation was detected by MTT assay, apoptosis was detected by Annexin V-FITC/PI double staining, proliferation and apoptosis-related protein expression were detected by Western blot. Results Triptolide inhibited the expression of CXCR4 mRNA and protein in A549 cells (P＜0.05). Triptolide inhibited the proliferation of A549 cells and induced apoptosis(P＜0.05). Transfection of pcDNA-CXCR4 up-regulated the expression of CXCR4 mRNA and protein (P＜0.05). Up-regulation of CXCR4 expression partially reversed the effect of triptolide on proliferation inhibition and apoptosis induction of A549 cells (P＜0.05). Conclusions Triptolide may inhibit the proliferation of non-small cell lung cancer A549 cells and induce apoptosis by down-regulating the expression of CXCR4.

miR-150-5p enhances radiotherapy sensitivity of hepatocellular carcinoma cells line HepG2 through targeting SIRT1

WU Da-ping, XU Lu, WU Huan-liang, ZHENG Wen-hong, ZHENG Xiao-mei, XIE Wen-rui

2020, 40(3):  321-327. 

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Objective To investigate the effect on the radiosensitivity enhancement of hepatocellular carcinoma cells by targeting SIRT1 with miR-150-5p. Methods The establishment of radioresistant cell strain RR-HepG2 was induced by fractional incremental radiotherapy. RT-qPCR was used to detect the expression of miR-150-5p in HepG2 and RR-HepG2 at different radiation doses. Cell cloning assay was used to detect the radiosensitivity of the two groups cell at the same radiation dose. Flow cytometry and Western blot were used to detect the effect of over-expression of miR-150-5p on apoptosis of HepG2 cells. Double luciferase assay was used to detect the association between miR-150-5p and SIRT1.Cell cloning and Western blot were used to detect the changes of radiosensitivity and apoptotic protein after over-expression of SIRT1. Results The expression of miR-150-5p in RR-HepG2 group decreased significantly with the same radiation dose(P＜0.05). The survival fraction and sensitivity of cells in agomiR-150-5p group decreased significantly(P＜0.05), the expression of Bax and caspase-9 increased significantly, the expression of Bcl-2 decreased significantly, and the apoptotic rate was significantly increased. The wild type (WT)agomiR-150-5p luciferase activity decreased significantly and SIRT1 protein level decreased significantly(P＜0.05); wild type (WT)anta-agomiR-150-5p luciferase activity increased significantly and SIRT1 protein level increased significantly (P＜0.05); WT-anta-agomiR-150-5p luciferase activity increased significantly and SIRT1 protein level increased significantly (P＜0.05). The survival fraction of cells in the agomiR-150-5p group decreased significantly, the expression of Bax and caspase-9 increased significantly, and the expression of Bcl-2 decreased significantly(P＜0.05). The survival fraction of cells in the agomiR-150-5p+SIRT1 group increased significantly, the expression levels of Bax and caspase-9 decreased significantly, and the expression of Bcl-2 increased significantly(P＜0.05). Conclusions The down-regulation of SIRT1 expression by miR-150-5p improves the radiosensitivity of hepatocellular carcinoma cells and provides a potential target for clinical radiotherapy of hepatocellular carcinoma.

Down-regulation of Notch1 inhibits proliferation and induces apoptosis of fibroblasts from pathological scars

SU Jiang-wei, YU Hui, DU Kun, WANG Jie, LIU Lin, CAI Jian

2020, 40(3):  328-333. 

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Objective To investigate the effect of down-regulation of Notch1 expression mediated by lentivirus on the activation of caspase-9 and mitochondrial membrane potential of fibroblast from pathological scars(PSF). Methods PSFs were isolated and cultured and then infected by Notch1 siRNA lentivirus and negative control lentivirus respectively.The level of Notch1 mRNA and protein was detected by real-time PCR and Western blot,respectively.MTT assay was used to detect cell proliferation; PI monochrome staining was used to detect cell cycle changes. Annexin V-FITC/PI assay was used to detect apoptotic changes; Western blot was used to detect the level of cyclin D1, CDK4, c-caspase-3, c-caspase-9 and cytochrome C protein levels in cytoplasm and mitochondria; JC-1 method was used to detect the mitochondrial membrane potential. Results The expression of Notch1 in PSFs transfected with Notch1 siRNA lentivirus was significantly lower than that in the negative control lentiviruses and the non-transfected PSFs(P＜0.05).The proliferation of PSFs decreased after Notch1 was down-regulated (P＜0.05);the percen-tage of G₀/G₁ phase increased(P＜0.05); the apoptotic rate increased(P＜0.05); the expression of c-caspase-3 and c-caspase-9 protein increased(P＜0.05); the mitochondrial membrane potential of the cells decreased significantly(P＜0.05); the level of cyclin D1 and CDK4 protein in cells decreased(P＜0.05); cytochrome C protein increased in cytoplasm and decreased in mitochondria(P＜0.05). Conclusions Down-regulation of Notch1 inhibits proliferation and induces apoptosis of PSF.

Propofol inhibits proliferation, migration and invasion of human lung adenocarcinoma cell lines through down-regulating PKM2

HU Zhi-hui, LI Jian-qin, HAN Lu-jun, ZHANG Jing, ZHANG Hong-xin

2020, 40(3):  334-339. 

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Objective To investigate the mechanism of propofol on proliferation, migration and invasion of lung adenocarcinoma cells. Methods MTT method was used to detect the inhibition rate or proliferation of A549 and Anip973 human lung adenocarcinoma cell lines treated with propofol (60, 100, 120 μmol/L); The si-NC group (transfected si-NC), the si-PKM2 group (transfected si-PKM2), pcDNA3.1 group (transfected pcDNA3.1), pcDNA3.1-PKM2 group (transfected pcDNA3.1-PKM2), propofol + pcDNA3.1 group (transfected pcDNA3.1 and treated with propofol), propofol + pcDNA3.1-PKM2 group (transfected pcDNA3.1-PKM2 and treated with propofol), all were transfected into Anip973 cells by liposome and treated with propofol; Transwell assay was used to detect cell migration and invasion; RT-qPCR was used to detect the expression of PKM2 mRNA in cells; Western blot was used to detect protein expression of PKM2, E-cadherin, MMP-2, in cells. Results Propofol(0, 60, 100, 120 μmol/L) inhibited the proliferation of human lung adenocarcinoma cells A549 and Anip973 in a concentration-dependent manner. Anip973 cells were more sensitive to propofol, and the optimal concentration was 120 μmol/L; propofol inhibited the proliferation, migration and invasion of Anip973 cells, down-regulated the protein expression of PKM2 and MMP-2, and up-regulated the protein expression of E-cadherin; knockdown of PKM2 has the same inhibition effect as propofol proliferation, migration and invasion, down-regulate the protein expression of MMP-2 and up-regulate the protein expression of E-cadherin of Anip973 cells; over-expression of PKM2 reversed the proliferation, migration and invasion, the effect of protein expression of E-cadherin and MMP-2 of Anip973 cells. Conclusions Propofol inhibits the proliferation, migration and invasion of lung adenocarcinoma cells, and its mechanism is related to down-regulation of PKM2 expression.

Taurine combined with SN50 inhibits the proliferation of human hepatocellular carcinoma cell line HepG2

LEI Qiao-li, RAO Qi-bin, LIN Ze-wen

2020, 40(3):  340-345. 

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Objective To investigate the effect of taurine combined with NF-κB signaling pathway inhibitor SN50 on the proliferation of hepatocellular carcinoma cells line HepG2 and its mechanism. Methods The expression of p-IκBα and IκBα proteins in hepatoma cells line HepG2 was detected by Western blot after the treatment with 150 mmol/L taurine for 24 hours. HepG2 cells were randomly divided into control group (untreated), taurine group (treated with 150 mmol/L taurine for 24 hours), inhibitor group (treated with 36 μmol/L SN50 for 24 hours) and taurine + inhibitor group (treated with 150 mmol/L taurine and 36 μmol/L SN50 for 24 hours), cell proliferation and apoptosis were examined by MTT, colony forming test and flow cytometry respectively,and cyclin D1, Bcl-2 protein and mRNA were detected by Western blot and RT-qPCR. Results After treatment with 150 mmol/L taurine for 24 hours, the expression of p-IκBα protein in HepG2 cells decreased in a time-dependent manner (P＜0.05). After treatment with 150 mmol/L taurine or 36 μmol/l SN50 for 24 hours, the survival rate and cloning rate of HepG2 cells significantly decreased, the apoptosis rate significantly increased, and the expressions of cyclin D1 and Bcl-2 protein and mRNA were significantly inhibited as compared with the control group, all the differences were statistically significant(P＜0.05).At the same time, SN50 enhanced the inhibitory effect of taurine on the proliferation of HepG2 cells, the expression of cyclin D1 and Bcl-2, and the promotion of apoptosis. Conclusions Combined efforts of taurine with NF-κB signaling pathway inhibitor SN50 inhibit strongly the proliferation of hepatoma cells line HepG2 through NF-κB signaling pathway.

Expression of vWF in transplanted human lung cancer with the different metastatic potential

SI Xiao-li, LI Shuang-yan, GUO Chao, ZHANG Song, ZHANG Li

2020, 40(3):  346-350. 

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Objective To explore the role of von Willebrand factor (hereinafter referred to as vWF) in the transplantation of human lung cancer with the different metastatic potential. Methods Low and high metastatic human lung cancer cell line A549 and D95 were cultured in vitro.The invasion and migration of lung cancer cells were detected by Transwell assay.Ten Balb/c nude mice were randomly divided into A549 group and D95 group(n=5).Then 0.2 mL of two tumor cell suspensions(the concentration was about 1×10⁷/mL) were subcutaneously injected respectively into the left upper limb of nude mice in A549 group and D95 group,to establish the xenograft model of human lung cancer in nude mice. The effects of vWF on transplantation tumor were analyzed.The level of vWF in peripheral blood was evaluated by ELISA.The expression of vWF protein in transplanted tumor was measured by immunohistochemistry.The expression of vWF protein in transplanted tumor, lung and liver was examined by Western blot. Results The number of invasion and migration of D95 lung cancer cells in vitro was more than that of A549 group(P＜0.05).After subcutaneous inoculation, all nude mice in the D95 group presented cancer nodules on the 5th day, and all nude mice in the A549 group did on the 7th day.Compared with the A549 group,the transplanted tumor mass and the quantity of lung metastatic tumor nodules in D95 group significantly increased(P＜0.05).Meanwhile,the expression of vWF protein in the transplanted tumor, lung and liver tissue were remarkably higher than that in the A549 group(P＜0.01). Conclusions vWF is closely related to the metastatic potential of human lung cancer cells.

Pyk2 promotes Ang Ⅱ-induced cardiac hypertrophy in rat

YAN Wen, LIU Li-xin, KONG Xiang-zhao, LUO Yu-chen, LI Zhong-qiu, LI Nan

2020, 40(3):  351-355. 

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Objective To investigate the role of proline-rich protein tyrosine kinase (Pyk2) in angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy and the underlying mechanism in rat. Methods Primary rat cardio-myocytes were isolated from rats and randomly divided into control group, Ang Ⅱ induction group (the dose of Ang Ⅱ was 100 nmol/L, cardiomyocytes were stimulated in the cell culture dish for 24 hours), Pyk2 inhibitor PF-431396 group (the dose of PF-431396 was 10 μmol/L, added to the cell culture dish 30 minutes in advance before Ang Ⅱ stimulation) and Ang Ⅱ induction group intervened by PF-431396 (the dose of Ang Ⅱ was 100 nmol/L, the dose of PF-431396 was 10 μmol/L). The size of cardiomyocytes was measured by α-actinin fluorescence staining. RT-qPCR was performed to detect the mRNA level of hypertrophic markers (ANP and β-MHC) in cardiomyocytes. The expressions of p-Pyk2, Pyk2 and p53 in cardiomyocytes were detected by Western blot analysis. Results Ang Ⅱ stimulation increased the surface area of cardiomyocytes and the mRNA level of hypertrophic markers(P＜0.05). In addition, the proteins of p-Pyk2 and p53 were increased(P＜0.05). However, after treatment of Pyk2 inhibitor PF-431396, moycardial hypertrophy was significantly improved and the protein level of Pyk2 and p53 decreased(P＜0.05). Conclusions Pyk2 promotes cardiomyocyte hypertrophy in rat induced by Ang Ⅱ through the p53 signaling pathway.

Myrtol standardized reduces the damage of PM2.5 exposure on mucosal cilia in rat airway

DU Ning, LU Xiao-qiang, DUAN Zheng

2020, 40(3):  356-362. 

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Objective To investigate the damage effect of fine particulate matter(PM2.5)exposure on mucociliary clearance, and analyze the effect of myrtol standardized on it. Methods Rats were divided into control group(group N),PM2.5 exposure group(group PM) and myrtol standardized group(group PM+CS).Rats were respectively given inhalation to filtered clean air indoor, exposure to ambient air of Shijiazhuang city (the range of PM2.5 concentration from 29.90 mg/m³ to 99.65 mg/m³) and gavage of myrtol standardized suspension 300 mg/(kg·d).Eight rats in each group were sacrificed after 3 months and 6 months.The lung and tracheal tissues were stained with HE and AB-PAS.The ultrastructure of cilia was observed by scanning electron microscope.The number of total cells,neutrophil count and classification in the BALF were detected by microscopy.The gene expression level of MUC5AC and AQP5 in pulmonary tissue were measured by real-time PCR. The protein expressional level of MUC5AC in lung tissue was respectively measured by immunohistochemistry and Western blot. Results In the PM groups, inflammatory injury and myxocyte metaplasia of lung and tracheal tissues were obvious, and tracheal cilia were adhesive, lodging and absent, sparse and disordered.The total number of white blood cells and percentage of neutrophils in BALF were significantly higher than those in N groups(P＜0.05).The expression of MUC5AC mRNA and MUC5AC protein in PM groups was higher than that in N groups(P＜0.05),but the relative expression of AQP5 mRNA in PM groups was significantly lower than that in N groups(P＜0.05).Compared with PM group, myrtol standardized can reduce the above damage effect(P＜0.05). Conclusions Myrtol standardized can protect the mucociliary clearance system in rats by reducing the damage of cilia structure, hypersecretion of mucus and inflammatory damage caused by PM2.5.

Electric stimulation enhances recovery of motor function in spinal cord contused rats

ZENG Xi, ZHAN Qi, GAO Yuan, CUI Nai-hao, FENG Lu-qian

2020, 40(3):  363-367. 

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Objective To investigate the effect of nerve electric stimulation on spinal cord contused rats and to explore the appropriate mechanism. Methods Rats were divided into sham group, control group and experimental group. T9 rat spinal cord contusion(SCC) model was developed by Allen's method. Experimental group was treated with nerve electric stimulation. After neurobehavioral score assessment, immunohistochemistry and Western blot were used to detect nerve growth factor and nestin. Results After spinal cord injury, score of rats gradually increased,and compared with control group, the experimental group significant increased(P＜0.05) at d 7. The nerve growth factor and nestin expression of experimental group, up-regulated significantly (P＜0.05). Conclusions Nerve electric stimulation may create a microenvironment conducive to neural survival and differentiation, which facilites the recovery of hindlimb motor function in rats.

rL-IL29 inhibits proliferation and migration of human gastric cancer cell line BGC

YIN Chao-yun, ZHANG Yao, CHEN Zhen-wei, ZHANG An-wei, ZHANG Xuan-feng, ZHANG Ri-ting, BU Xue-feng

2020, 40(3):  368-373. 

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Objective To investigate the effect of recombinant Newcastle disease virus expressing IL-29(rL-IL29) on the proliferation and migration with invasion of gastric cancer cells and to explore the potential mechanism. Methods Gastric cancer cell lines BGC were cultured and divided into the PBS, rL-IL29 and newcastle disease virus(NDV)groups infected by the PBS, rL-IL29 or NDV.Western blot was used to detecte the expression of NDV, IL-29,p-ERK,MMP2 and p-AKT protein in cells.The proliferation,migration and invasion of gastric cancer cell lines BGC were detected by CCK8、cell clone forming experiments、scratch test and Transwell invasion test. Results IL-29 protein was only expressed in rL-IL29 group,NDV protein were expressed both in rL-IL29 group and NDV group and was significantly higher than those of PBS group (P＜0.05). The proliferation and migration of BGC cells infected with rL-IL29 were inhibited more seriously than those infected with NDV or PBS group.The expression of p-ERK,MMP2 (66 ku/72 ku),p-AKT and P65 protein was all significantly decreased(P＜0.05). Conclusions The proliferation and migration of gastric cancer cell lines BGC were inhibited by rL-IL29 and the mechanism may be related with ERK,AKT signaling pathways.

Dexmedetomidine relieves ischaemia-reperfusion injury of kidney via up-regulation of sirtuin 3

WU Yue-ling, QU Hai-xia, WANG Jun-xing

2020, 40(3):  374-379. 

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Objective To study the protective effect and mechanism of dexmedetomidine (DEX) on renal ischemia-reperfusion (I/R) injury in mice. Methods The mice were divided into sham group, siRNA+sham group, renal I/R group (conventional method), DEX+I/R group (25 μg/kg DEX, intraperitoneal injection), siRNA+I/R group and siRNA+DEX+I/R group; siRNA+sham group, siRNA+I/R group and siRNA+DEX+I/R group were injected 5×10⁹ TU/kg lentivirus-sirtuin3 (SIRT3) siRNA through tail vein 3 days before surgery. Renal function was measured by biochemical analyzer, renal histomorphology was observed by HE staining, renal tissue apoptosis was detected by Tunel staining, and the expression of SIRT3, cyclophilin D (Cyp D) and cytochrome C (Cyt C) were detected by Western blot. Results DEX reduced the renal pathological damage after I/R, up-regulate the expression of SIRT3 and down-regulated the expression of Cyt C and acetylated Cyp D (P＜0.05). Inhibition of SIRT3 attenuated the protective effect of DEX on kidney. Conclusions The role of DEX in alleviating renal I/R injury may be related to the up-regulation of SIRT3.

Shikonin attenuates hypoxia-reoxygenation-induced injury of cardiomyocyte cell line H9c2

LIU Ming, YANG Chen-li, MENG Qing-xin

2020, 40(3):  380-387. 

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Objective To explore the protective effect of shikonin on H9c2 cardiomyocytes damaged by hypoxia-reoxygenation(H/R). Methods H9c2 cells were divided into control group, H/R group, shikonin (0.1, 1 and 10 μmol/L) group, including 3 parallel wells in each group; MTT assay was used to detect the toxicity of shikonin on H9c2 cells; Apoptosis and cell cycle were detected by flow cytometry; DCFH-DA was used to detect the level of reactive oxygen species; biochemical detection of MDA and 8-OHdG in cells And GSH content; qPCR was used to detect the expression of γ-GCS, HO-1 and NQO1 mRNA; Western blot was used to detect the expression of Bax, Bcl-2, caspase-3, cleaved caspase-3, Keap1 and Nrf2 proteins; immunofluorescence was used to detect Nrf2 cells. Protein into the nuclear situation. Results Shikonin inhibited the proliferation of H9c2 cells in a concentration-time-dependent manner (P＜0.05). H/R induced apoptosis of H9c2 cells, cell cycle arrest and promoted the expression of Bax and cleaved caspase-3 proteins, inhibited the expression of Bcl-2 protein (P＜0.05). Shikonin inhibited apoptosis, relieved cell cycle arrest, and cleaved caspase-3 and Bax protein expression, and increased Bcl-2 protein expression in a concentration-time-dependent manner; Shikonin decreased the active oxygen, MDA and 8-OHdG induced by H/R, increased the GSH content induced by H/R and down-regulated the expression of γ-GCS, HO-1 and NQO1 mRNA, caused an increase in Nrf2 protein and down-regulation of Keap1, and promoted Nrf2 protein entry into the nucleus (P＜0.05). Conclusions Shikonin attenuates myocardial cell injury induced by hypoxia-reoxygenation in H9c2 cells.

Clinical Sciences

Margins of planning target volume in pancreatic cancer with tomotherapy

HE Lei, SUN Xian-song, YU Lang, WANG Xin-hai, HU Ke, QIU Jie, ZHANG Fu-quan

2020, 40(3):  388-391. 

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Objective Analysis of the pendulum error of pancreatic cancer patients by spiral tomography technique under the guidance of megavolt CT (megavoltage computed tomography, MVCT) image, and calculation of planned target volume, PTV) of the margin. Methods The MVCT image scan matched with the planned CT image, the error values of the left and right (X), head and feet (Y), abdominal back (Z) axis direction and cross-section rotation (roll) direction were recorded, and the error values were analyzed and calculated. Results A total of 592 MVCT scans were conducted. The X, Y, Z and roll direction swing error values were (-0.5±2.8)mm, (-1.1±6.4)mm, (6.0±4.4)mm, and (-0.2±0.7) respectively. The average error in the three directions of X, Y and Z was less than 5 mm, with 97.13% (575/592), 88.01% (521/592) and 37.84% (224/592), and roll direction rotation error of less than 1 degree, with 93.92% (556/592). According to the formula, the extra-boundary spacing values of The PTV in X, Y and Z 3 directions were 5.16, 9.9 and 7.53 mm respectively. Conclusions In order to improve the accuracy and efficacy of tomotherapy, The margin emission values of CTV to PTV in X, Y and Z three-dimensional directions are recommended as 5, 10 and 8 mm, respectively.

Mini Reviews

Research progress on the role of calcium sensing receptor in atherosclerosis

SHANG Yu-jia, LI Yu-xia, SONG Jia-xin, LI Hong-xia

2020, 40(3):  395-398. 

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Calcium sensing receptor (CaSR) is a member of the G protein-coupled receptor C family.It is widely expressed in different tissue cells in the body and functions in various pathophysiological processes of the body. CaSR can participate in the development of atherosclerosis by regulating vascular endothelial cell injury,inflammatory response,proliferation and migration of vascular smooth muscle cells and vascular calcification.

Research advances on biological functions of galectin-7

WANG Ran-ran, JIANG Shuang-quan, LI Shou-qiang, YU Dan-dan, LENG Xiao-ping

2020, 40(3):  399-402. 

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Galectins construct a family of animal lectins for β-galactosides, which are widely distributed from living nematodes to mammals and other living organisms,participating in various biological processes, such as cell-cell adhesion, growth regulation and immune response.Galectin-7, one of the most important member of the family is closely related to the differentiation and maturation of epithelial tissues, the proliferation of injured epidermis, the development of tumors and the immunity of body.

Research progress of immune checkpoint inhibitor related autoimmune hypophysitis

LI Jia-yi, XING Bing

2020, 40(3):  403-406. 

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Immunotherapy induced hypophysitis (IH) has been demonstrated an autoimmune adverse effect of immune checkpoint inhibitors treatment of malignant tumors. The incidence of IH depends on types of immunotherapeutic agents. Patients with IH often present non-specific symptoms, ACTH, TSH and gonadotrophin deficiencies, and radiographic pituitary enlargement. Hormone replacement is recommended to be applied on IH patients. High-dose glucocorticoids but not supported by recent studies. This study systematically reviewed the prevalence, epidemiological characteristics, clinical characteristics, treatment and prognosis of IH.

Medical Education

Application of 3D printing knee joint model in osteoarthritis clinic teaching for eight-year program medical students

NIU Shun, MA Bao-an, LONG Hua, WANG Dan, CHEN Hui-yun, ZHANG Yan, ZHOU Yong

2020, 40(3):  407-410. 

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Objective To explore the teaching effect of 3D printing knee joint model in eight-year osteoarthritis lecture. Methods Eight-year program students in grade 2014 were selected and randomly divided into an experimental group and a control group. 3D printing knee joint model display + animation demonstration + LBL teaching were used in the experimental group, while animation demonstration + LBL teaching were used in the control group. Teaching effect was evaluated through examination scores and teaching quality evaluation. Results The examina-tion scores, teaching quality scores and the excellent rate of teaching quality were all significantly higher in the experiment group than those in the control group (P＜0.05). Conclusions The 3D printing model can enhance the learning enthusiasm and improve students' mastery of knowledge and so is recommended.

Application of three-dimensional scanning in rebuilding female reproductive system model for embryo transfer training

RAO Jin-peng, QIU Feng, CAI Yi-ting, WEI Kai, JIN Min, JIN Fan

2020, 40(3):  411-414. 

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Objective To explore three-dimensional scanning and reconstruction technology in embryo transfer training. Methods Two sets of female reproductive system models, standard and cervical stenosis, were reconstructed with three-dimensional scanner, data processing software EXScan-Pro and 3D printer. Twenty clinical and laboratory personnel were trained to simulate the transfer of degenerated embryos, and the accuracy of embryo implantation site, the time required for embryo transfer and the frequency of replacing the transfer catheter before and after the training were evaluated. Results After training, the accuracy score of embryo implantation site was increased from 0.60±0.41 to 1.37±0.29 (P＜0.05); the time required for embryo transfer was shortened from (340±146) s to (180±59) s (P＜0.05); the frequency of replacing the transfer catheter was decreased from (0.83±0.37) to (0.23±0.13) times (P＜0.05). Conclusions Application of rebuilding female reproductive system model for embryo transfer training can improve the practical experience of operators prior to formal surgery, improve the stability of embryo transfer and help to establish a standardized operation process, as a supplement to the traditional observation and teaching methods.

An exploration on OBE-based teaching model of medical metabolic biochemistry

YANG Rong, HUANG Jian, TONG Xue-mei, ZHANG Ping, YAO Yan-hua, WU Li-fang

2020, 40(3):  415-418. 

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Outcome-based education (OBE) is a kind of advanced education concept, is becoming the mainstream concept of medical education reform. Since “Medical Metabolic Biochemistry” integration module has only a few teaching hours, how to improve the quality of teaching and stimulate students' interest in learning is a real challenge. Based on the OBE education concept, this article aims to summarize the teaching reforms related to learning outcomes, teaching contents and teaching strategies, in order to enhance students' enthusiasm and initiative in learning, so as to improve teaching effectiveness, and also to strengthen the foundation for students to become medical talents that meet the requirements of “Outstanding Talent Education Training Scheme 2.0”.

Remodeling of clinical microbiology teaching mode

HUANG Jing-jing, SUN Hong-li, FAN Hong-wei, XIE Xiu-li, ZHANG Zhan-jie, XU Ying-chun

2020, 40(3):  419-422. 

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Laboratory Diagnosis of Infectious Diseases and Hospital Infection Control by Common Pathogens, one of the curriculum of Laboratory Diagnostics is both a starting point for most medical students to understand clinical microbes, and is also a platform for them to learn relevant basic knowledge. In the past, the course was instilled by teachers through electronic courseware in the classroom.As a result, student's learning passion is hard to be stimulated whtn lacked practical and intuitive understanding. In order to give full play to students' subjective initiative and maximize their interest in learning, the teaching team havemade new attempts and explorations on the form of teaching. The new mode enhances the capacity of self-learning, teamwork, and presentation to communication by increasing group interactive discussions, and presents a new look of learning and immersive learning with physical teaching.

Application of lean management in the optimization of thyroid puncture appointment process

LI Guang-han, LI Ying, GUO Ke-ke, GAO Peng, ZHANG Wei-shuo, TU Ling, ZHANG Bo

2020, 40(3):  423-428. 

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Objective To optimize the appointment process of thyroid fine needle aspiration by using the theory of lean management. Methods From June to September 2019, the patients who had thyroid puncture appointment in the ultrasound department of China-Japan friendship hospital were studied.Fifty patients before and after the process improvement were randomly included in the control group and 50 patients after the process improvement were in the study group. A total of 100 patients were collected. The total time of the appointment process, the total walking distance and the number of round trips of the appointment process, and the patient satisfaction were compared between the two groups. Results After the improvement, the time of appointment process was reduced from (58.6±31.5)min to (32.5±12.8)min; the satisfaction score of patients was increased from 3.3 to 4.3; and the walking distance was reduced from (90~546)m to (94~219)m, among which the walking distance involving the application form and payment of ultrasound examination was reduced from 78-149 m to 7 m, the total number of waiting in queues reduced from 3-7 to 1-2. Conclusions Based on the optimization method of lean management, it can optimize the process of patients' appointment and improve patients' satisfaction, which is conducive to the more efficient operation of the hospital's overall resources.

From the perspective of integrative medicine to help western medicine learners understand the theory of viscera and bowels

QU Ling, LIANG Xiao-chun

2020, 40(3):  429-432. 

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Traditional Chinese medicine(TCM) theory of viscera and bowels is as anatomy, physiology, pathophysiology, but they have essential differences. Students in western medicine colleges frequently get confused when they are learning this part of the course. The fundamental cause of the confusion is that two different medical theoretical system of Chinese and western describe the same organ in human body and the result of the paper summarizes the phenomenon of life from different angles, using different methods and frame. Therefore, the scientific and reasonable combination of modern medical knowledge to explain the functions of TCM viscera and bowels will help students in western medical colleges to better understand relevant knowledge.