Yin Yuan , Xu Wei , Wu Shuangshuang , Li Yan , Zhao Weihong , Wu Jianqing
2017(9):1067-1075. DOI: 10.7655/NYDXBNS20170901
Abstract:Objective: To investigate the role of miRNA-200a in the efficacy of targeted therapy in non-small-cell lung cancer (NSCLC) patients and its underlying mechanisms. Methods: Real-time polymerase chain reaction and Western blot were used to investigate the level of miRNA-200a and FOXC1. The MTT assay,wound-healing and Transwell assays were performed to measure the effect of miRNA-200a and FOXC1 on cell growth, migration, invasion and other biological behaviors. Luciferase reporter assay analyzed the relationship between FOXC1 and miRNA-200a. Results: We found that a high level of miRNA-200a inhibited NSCLC cells growth, EMT, migration and invasion and increased sensitivity to gefitinib by targeting FOXC1. Furthermore, suppression of FOXC1 also inhibits cells progression and restores gefitinib resistance. Conclusion: Upregulated miRNA-200a or knockdown of FOXC1 enhanced sensitivity to gefitinib in NSCLCs. This may provide a novel effective therapeutic approach to overcome the acquisition of resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) therapy.
Han Guoyong , Zhang Long , Chen Zhiqiang , Wang Xiaowei , Zhu Qin , Huang Xinli , Wang Xuehao
2017(9):1076-1080. DOI: 10.7655/NYDXBNS20170902
Abstract:Objective: To investigate the expression of miRNA-873 in hepatocellular carcinoma(HCC) tissues and cell lines, further study the effect of miRNA-873 on migration and invasion in hepatocellular carcinoma cell lines. Methods: The expression of miRNA-873 in HCC tissues and cell lines was detected by RT-PCR. Wound-healing experiment and Transwell invasion were used to examine migration and invasion of cells, respectively. Western blot was used to detect the activity of Janus kinase 2(JAK2) and signal transduction and transcriptional activation 3(STAT3). Results: The results showed that miRNA-873 was upregulated in HCC tissues and HCC cell lines, compared with that in the normal control. RT-PCR showed that the expression of miRNA-873 decreased or increased after miRNA-873 inhibitor or mimic transfection, respectively(P<0.01). Compared with the normal control, knockdown of miR-873 significantly inhibited the migration and invasion of HepG2(P<0.05). However, overexpression of miRNA-873 had the opposite effect(P<0.05). We also found that miRNA-873 significantly activated JAK2/STAT3 signal pathway. Conclusion: MiRNA-873 was overexpressed in HCC and might promote HCC migration and invasion through JAK2/STAT3 signal pathway.
Kong Chengcheng , Yang Wenping , Hua Jian , Liu Yan , Ding Haixia
2017(9):1081-1085. DOI: 10.7655/NYDXBNS20170903
Abstract:Objective:To investigate the neuroprotective mechanism of serum miRNA-181a on dopaminergic neurons in primary cell model of Parkinson’s disease induced by 1-methyl-4-phenyl-pyridinium (MPP+). Methods:Embryonic mouse mesencephalic primary neurons were cultured in vitro and divided into four groups:a blank control group, a MPP+ group, a miRNA-NC+MPP+ group, a miRNA-181a+MPP+ group. Dopaminergic neurons dendrites and axons were identified and counted by using tyrosine hydroxlase(TH) immunostaining under microscope in each group. Results:The dopamin(DA) neurons exhibited intact circular or triangular bodies and a variety of dendrites and axons derived from bodies, and had intact morphous and interweaved closely in the control group. The group treated by MPP+ were specifically found that the number of dopaminergic neurons decreased strikingly, axons and dendrites disappeared completely, and bodies remained. The axons appeared abnormal varicosities, apparent string-of-beads change. The axons and dendrites almost fragmented, only few cell bodies disappeared and exhibited axonal root. In the MPP+ group, the number of dopaminergic neurons increased, and cell bodies, axons, dendrites and the death of the dopaminergic neurons became improvement. Conclusion: The results suggested that miRNA-181a may play an important role in protecting dopaminergic neurons in MPP+ induced Parkinson’s disease.
Zheng Rui , Du Junjie , He Keshuai , Shao Yongfeng
2017(9):1086-1092. DOI: 10.7655/NYDXBNS20170904
Abstract:Objective: To investigate the calcification mechanism of bicuspid aortic valve(BAV) which involves the microRNA-195. Methods: Stenotic BAV or degenerative tricuspid aortic valve(DTV) samples were collected from 35 patients undergoing aortic valve replacement. The mRNA expression of miRNA-195 and SMAD7, and protein expression of SMAD7 were determined by RT-PCR and Western blot, respectively. Dual-luciferase assay was performed to determine putative target of miRNA-195. In porcine valve interstitial cells(VIC) ,we down-regulated and up-regulated the microRNA-195 level, and examine the mRNA expression of SMAD7 to explore the relationship between miRNA-195 and SMAD7. Moreover, in human leaflet samples, immunochemistry staining was carried out to test the protein expression of MMP-2 and MMP-9, and Sirus Red staining was done to investigate the expression of collagen, and the Von Kossa staining was implemented to explore the mineralization nodes. All of these were done to compare the functional alteration associated with cellular calcification. Results: Compared with TAV, the expression of miRNA-195 was remarkably lower in BAV with higher expression of SMAD7. Luciferase experiments validated that miRNA-195 directly targeted SMAD7. Overexpression of miRNA-195 inhibited the mRNA expression of SMAD7, while miRNA-195 inhibitor treatment resulted in increased SMAD7 expression in VIC. Finally, more MMP-2 and MMP-9 expression and more collagen distribution were observed in BAV than in DTV. Conclusion: Our study has demonstrated that miRNA-195 is much more down-regulated in stenotic BAV than that in DTV. The downregulation of miRNA-195 promotes valvular calcification via targeting SMAD7, which involves the fibrosis of extracellular matrix.
Guo Xin , Cai Yunlang , Ren Mulan
2017(9):1093-1098. DOI: 10.7655/NYDXBNS20170905
Abstract:Objective:We sought to construct hollow mesoporous silica nanoparticles(HMSN) compound drug delivery system, which can target ovarian cancer cells, and to assess the effect of this drug delivery system as well as explore the treatment effect of insulin-like growth factor receptor(IGF-1R) specific inhibitor NVP combined with doxorubicin(DOX) by HMSN delivery system. Methods: We modified carboxyl on the surface of HMSN, then loaded DOX and fluorescent NVP through mechanical mixing and the electrostatic attraction. The compound drug system was characterized by TEM, infrared spectrometer and Zeta potential. The drug release rates of DOX and fluorescence NVP were detected in different pH environment. Drug delivery system gathered in cells was observed by immunofluorescence laser scanning confocal microscope at different times. We divided this experiment into HMSN-DOX-NVP group, HMSN-DOX group and the control group, and acted on SKOV-3 cells respectively. Cell apoptosis rates and cell inhibition rates in there three groups were detected by flow cytometry. Results: Zeta potentially showed that the charge of HMSN was about -22.3 mV before modified carboxyl, and the charge of HMSN raised to -44.9 mV after modified carboxyl. The size and shape of HMSN were not changed after loaded drugs observed by TEM. The drug release rates of DOX and fluorescence NVP were gradually increased with the environmental pH value gradually reduced. HMSN-DOX-NVP and HMSN-DOX gathered in cells and gradually infiltrated into the nucleus with the prolongation of time. The cell apoptosis rate and MTT inhibition rate of the HMSN-DOX-NVP group were higher than those of the HMSN-DOX group and the free control group(P<0.05). Conclusion: HMSN compound drug delivery system can be well gathered in cytoplasm mainly through nonspecific endocytosis and ensure pharmacological activity of the released drugs simultaneously. NVP combine with DOX can improve the killing rate of DOX for cancer cells and achieve targeted inhibition of IGF-R.
Zhou Xujie , Wu Jing , Zhu Lei , Sun Xi , Ding Qiang
2017(9):1099-1103. DOI: 10.7655/NYDXBNS20170906
Abstract:Objective: To investigate the effects of RNPC1 on the sensitivity of breast cancer cell MCF-7 with doxorubicin (DOX). Methods: Lentivirus was used to over-express and knock-down RNPC1 in the MCF-7 breast cancer cells. The cells were divided into overexpress RNPC1(RNPC1) group and its control(NC group), knock-down RNPC1(shRNPC1 group) and its control(SCR group). The relative mRNA and protein expression of RNPC1 was detected by qRT-PCR and Western blot, respectively. The experimental groups were treated with different concentration of DOX, and cell growth inhibition rate was tested cell counting kit 8(CCK-8). After the experimental groups was treated with DOX, cell apoptosis rate was analysis by flow cytometer assay and the expression of apoptosis-related protein was detected by Western blot assay. Results: The qRT-PCR and Western blot results showed that the mRNA and protein level of RNPC1 was increased after transfection with RNPC1 overexpression(RNPC1). While, RNPC1 was reduced after transfection with knockdown RNPC1 (shRNPC1) lentivirus. After treatment with different concentration of DOX, the IC50 of shRNPC1 group was significantly lower than SCR group (P <0.05). Furthermore, the IC50 of RNPC1 group was higher than NC group (P<0.05). The flow cytometer assay was used after treatment with DOX for 24 h in the experimental groups. The cell apoptosis rate of shRNPC1 group was higher than SCR group, while, the cell apoptosis rate of RNPC1 was lower than NC group. Overexpression of RNPC1 increased the expression of BCL-XL and BCL-2 after treatment with DOX. Reversely, knockdown of RNPC1 reduced the expression of BCL-XL and BCL-2. Furthermore, overexpression of RNPC1 reduced the expression of BIM and BID after treatment with DOX. While, knockdown of RNPC1 increased the expression of BIM and BID. Conclusion: RNPC1 could decrease the sensitivity of breast cancer cell MCF-7 to DOX, and the underlying mechanism might be related to cell apoptosis.
Guo Song , Peng Yunpeng , Lu Zipeng , Fu Yue , Yin Lingdi , Miao Yi
2017(9):1104-1108. DOI: 10.7655/NYDXBNS20170907
Abstract:Objective:To explore the effect of insulin on the pancreatic cancer cell lines and the potential mechanisms. Methods: The expression of insulin receptor(IR) was evaluated by qRT-PCR and Western blot. We examined the effect of insulin with different concentrations on the viability of pancreatic cancer cell lines to choose optimal concentration. After insulin treatment, the viability of pancreatic cancer cells was measured by CCK-8 assay; Invasion and migration of cells were measured by a Transwell chamber assay; Western blot was used to observe the expression of matrix metalloprotease(MMP)-2, MMP-7 and MMP-9. Results: Pancreatic cancer cell lines PANC-1 and Miapaca-2 showed relatively higher expression of IR(P<0.05). The cell lines had significantly enhanced viability(P<0.05) when treated with insulin of 100 nmol/L. CCK-8 assay showed that the proliferation of the cell lines was increased significantly. Transwell chamber assay showed that insulin significantly enhanced the invasion and migration of pancreatic cancer cells(P<0.05). Western blot proved that the expression of MMP-2, 7 and 9 was remarkably increased. Conclusion: Insulin can enhance the invasion and migration of pancreatic cancer cell lines by up-regulating MMP-2, 7 and 9.
Wang Ya’ni , Zhao Pengfei , Tan Wenfeng , Zhang Miaojia
2017(9):1109-1113,1153. DOI: 10.7655/NYDXBNS20170908
Abstract:Objective: To explore the effects of adiponectin receptor 1(AdipoR1) knockdown on the proliferation and apoptosis of human rheumatoid arthritis synovial fibroblast cell line(MH7A) induced by lipopolysaccharide(LPS). Methods: ①The interference efficiency of AdipoR1 silenced (sh-AdipoR1) MH7A cells was identified by immunofluorescence and Western blot techniques. ②The effects of LPS(100 ng/mL) on the proliferation and apoptosis of sh-AdipoR1 and sh-NC cells were detected by CCK8 kit and flow cytometry, respectively. ③ Besides, the expression levels of apoptosis associated genes: BCL-2, BCL-XL, BAX and BAK in sh-AdipoR1 and sh-NC cell lines stimulated by LPS, were detected by real-time polymerase chain reaction(real-time PCR) experiment. Groups of sh-NC and sh-AdipoR1 without LPS stimulation were set as controls. Results: ①The expression of fluorescent protein in MH7A cells transfected with lentivirus was nearly 100%, detected by fluorescence microscope. Moreover, the expression of AdipoR1 protein in sh-AdipoR1 group was significantly lower than that in sh-NC group(P<0.05), indicating that AdipoR1 silenced cells were successfully constructed. ②The cell proliferation rate and apoptotic cell rate of sh-AdipoR1 group had no obvious difference compared to sh-NC group without LPS stimulation. The cell proliferation rate of both sh-NC and sh-AdipoR1 groups increased after 24 and 48 hours LPS stimulation, and apoptotic cell rate of sh-NC and sh-AdipoR1 groups significantly increased after 48 hours LPS stimulation(P<0.05). The cell proliferation rate of sh-AdipoR1 group was significantly lower than that of sh-NC group after LPS stimulation for 24 and 48 hours (P<0.05). And the apoptotic cell rate of sh-AdipoR1 group was significantly higher than that of sh-AdipoR1 group after LPS stimulation for 48 hours(P<0.05). ③The mRNA expression levels of BCL-2 and BCL-XL of sh-AdipoR1 group had no significant difference compard to the sh-NC group without LPS stimulation. Neither did BAK and BAX mRNA expression levels. After LPS stimulation, BCL-2 and BCL-XL mRNA expression levels of both sh-NC and sh-AdipoR1 groups reduced, while BAK and BAX elevated(P<0.05). Compared with those in sh-NC group, the mRNA expression levels of BCL-2 and BCL-XL were significantly decreased while BAX and BAK were significantly increased in the sh-AdipoR1 group with LPS stimulation(P<0.05). Conclusion: Our research found that down-regulation of AdipoR1 gene could significantly inhibit the proliferation and promote the apoptosis of MH7A cells under LPS exposure. It indicates that adiponectin receptor-1 related signaling pathways may play a role in rheumatoid arthritis.
Zhao Ling , Yang Xiaofan , Zhou Xiaobin , Ji Xiaohui
2017(9):1114-1119. DOI: 10.7655/NYDXBNS20170909
Abstract:Objective: To investigate the level of IL-10 expressing on induced Treg cells(iTreg) such as CD4+CD25+T cells of SLE patients. Methods: Peripheral blood mononuclear cells(PBMCs) were isolated from healthy donors and SLE patients, respectively, and CD4+CD25-T cells were further isolated. Then the cells were induced and transformed by TGF-β in vitro; expression of intracellular and membrane IL-10 on CD25+ and CD25-,Foxp3+ and Foxp3- subsets of induced and transformed T cells were analyzed by flow cytometry. Results: Expression level of the intracellular IL-10(iIL-10) in CD25+ T cell subsets was obviously increased in both of the induced and transformed CD4+T cells from the normal controls and SLE patients, but it was significantly higher in the SLE patients. In the induced and transformed CD4+T cells both from the normal controls and SLE patients, expession level of the membrane IL-10 in CD25+ T cell subsets was obviously increased, while it had no difference between the SLE patients and the control group. In the normal subjects and SLE patients, the expession level of intracellular IL-10 in the induced and transformed CD4+CD25+ Foxp3+T cells subsets was obviously increased, and was significantly higher in the SLE patients than that of the control group. The expession level of membrane IL-10 was increased in the induced and transformed CD4+CD25+ Foxp3+T cells subsets, while no difference was observed between normal subjects and SLE patients. All the changes mentioned before have no correlation with SLEDAI. Conclusion: The IL-10 expression level was closely correlated with the induced and transformed CD4+CD25+ Foxp3+ Treg cells from the normal subjects. Abnormal expression of intracellular IL-10 was observed on the induced and transformed Treg cells from the CD4+CD25-T cell of SLE patients. The intracellular IL-10 exprssion level of CD4+CD25+Foxp3+ Treg cells from the SLE patients was significantly higher than that of the control group. And the intracellular IL-10 exprssion level of CD4+CD25+ Foxp3- Treg cells and CD4+CD25-T cell without induction was also significantly higher than that of the control group; But the expessions level of membrane IL-10 was not different between the SLE patients and the normal controls.
Ding Ling , Shi Huimin , Zhang Aiqing , Gan Weihua
2017(9):1120-1123. DOI: 10.7655/NYDXBNS20170910
Abstract:Objective:To analyze the effects of bufalin on autophagy induced by gentamicin(GM) in rat renal tubular epithelial cells (NRK52e) in vitro. Methods:NRK52e cells were cultured in vitro and divided into 4 groups: the control group, the GM group (2 mg/mL), the bufalin group(1×10-8 mol/L) and the GM+bufalin group(GM 2 mg/mL+bufalin 1×10-8 mol/L). The autophagy of NRK52e cells was observed by transmission electron microscopy. Effects of GM and bufalin on the expression of autophagy-related protein LC3Ⅱ and P62, and kidney injury molecule-1(KIM1) protein were examined by Western blot. Results:Transmission electron microscopy showed that the number of autophagosomes in the GM group was significantly increased compared with the control group, and the number of autophagosomes of the bufalin group was less than that in the GM group. Western blot showed that the relative expressions of LC3Ⅱ, P62 and KIM1 protein were significantly increased in the GM group compared with the control group (all P<0.05), however, the relative expressions of LC3Ⅱ, P62 and KIM1 protein were decreased in the bufalin group than those in the GM group (P<0.05). Conclusion:Gentamicin could activate the autophagy of NRK52e cells and up-regulate the expression of KIM1 protein, while bufalin could partially inhibit GM-induced NRK52e cell’s autophagy and the up-regulated expression of KIM1 protein. It may be the mechanisms by which bufalin acts the renoprotection.
Xu Chaoqi , Xu Yuyu , Jiang Pan , Feng Qing
2017(9):1124-1130. DOI: 10.7655/NYDXBNS20170911
Abstract:Objective:To study the effect of oleuropein in combination of acrolein on proliferation and apoptosis related to endoplasmic reticulum stress(ERS) in human bronchial epithelial cells. Methods:HBE cells were treated with oleuropein and acrolein. MTT assay was performed to detect cell viability. Plate clone formation assay was used to get colony forming efficiency. Hoechst 33258 staining and flow cytometric assay were applied to investigate the apoptosis of cells. The levels of ERS biomarkers including glucose-regulated protein 78(GRP78) and C/EBP homologous protein(CHOP) were measured by Western blot. The mRNA expressions of GRP78 and CHOP were analyzed by real-time PCR. Twelve male Sprague-Dawley rats were treated with acrolein alone through intraperitoneal injection, or olive leaf extract in combination through gavage. Lung tissues were obtained from the rats for detecting the expressions of GRP78 and CHOP. Results:HBE cell proliferation was inhibited by acrolein in a dose dependent manner. Cell prolife-
Cheng Jie , Jiang Lei , Huan Fei , Huang Yumeng , Wang Jun
2017(9):1131-1135,1181. DOI: 10.7655/NYDXBNS20170912
Abstract:Objective:To explore the effects of cell injury and the expression of inflammation related genes in rat microglial cells induced by methamphetamine(Meth). Methods:Cell viability and apoptosis induced by Meth were detected by MTT and in situ end labeling(TUNEL) after primary culture of SD fetal rat microglial cells.The effect of Meth on the expression of inflammatory cytokines mRNA in the microglial cells was evaluated by real time fluorescence quantitative polymerase chain reaction(real-time PCR).The secretion of interleukin(IL)-6,tumor necrosis factor(TNF)-α and inducible nitric oxide synthase(iNOS)in microglia cells induced by Meth were detected by ELISA method. Results:MTT showed that Meth reduced microglia viability in a concentration dependent manner(25,50,100,and 200 μmol/L) with statistically significant at a concentration of 200 μmol/L(P<0.05). TUNEL results showed that 200 μmol/L Meth significantly contributed to cell apoptosis compared with the control group(P<0.05). Q-PCR results showed that Meth reduced the expression levels of IL-24 and nitric oxide synthase 3(NOS3) in microglial cells for 24 hours. However,the expression levels of Peli3,Sigma receptor 1(sig1)-R,IL-1,IL-6 and Toll like receptor 4(TLR4) were up-regulated. In addition,protein levels were also confirmed that Meth promoted the secretion of IL-6 and TNF-α. Conclusion:Meth exposure obviously contributes to microglial damage. Moreover,it causes IL-1β,IL-1R,IL-6,TLR4 and other inflammatory factors mRNA expression changes,and promotes the secretion of IL-6 and TNF-α,which may damage the central nervous system,resulting in neurotoxicity.
Wu Zeyu , Shi Qiqi , Huanghe Shuyuan , Yang Lei , Lu Mengting , Mo Xuming
2017(9):1136-1141. DOI: 10.7655/NYDXBNS20170913
Abstract:Objective: To investigate whether succinic acid could exacerbate the early interstitial fibrosis of cardio-pulmonary arteries induced by monocrotaline in a rat model of pulmonary hypertension, and to explore its pathological and molecular mechanisms. Methods: A total of 30 healthy male SD rats aged 4 to 5 weeks and weighted 120~180 g were randomly divided into the control group(C group), the monocrotaline group(MCT group), and the monocrotaline and succinic acid group(MCT+SUC group), with 10 rats in each group. The MCT group and the MCT+SUC group were subjected to intraperitoneal injection with monocrotaline in 60 mg/kg. The MCT+SUC group was given SUC for 4 weeks. The rats in the control group and the MCT group were injected with equal volume of saline, lasting for 4 weeks. After 4 weeks, hemodynamic indices including mean pressure of pulmonary artery(mPAP), right ventricular systolic pressure(RVSP), systolic blood pressure(SBP) and diastolic blood pressure(DBP) of pulmonary artery were measured by floating catheter in all groups. The right ventricular hypertrophy index was measured after euthanasia. HE staining, Masson staining and Sirius red staining were conducted to detect the remodeling of arteriole in the heart and lung tissues. Western blot and immunohistochemistry were conducted to examine the protein level of angiotensinⅡ(AngⅡ), platelet derived growth factor(PDGF-BB), and platelet derived growth factor(FGF2) in the pulmonary hypertension rat model. Results: The rats in the MCT group and the SUC group both showed decreased activity, sleepiness and poor appetite. Lips cyanosis and shortness of breath appeared earlier in the MCT+SUC group. Hemodynamic examination exhibited that indices, including mPAP, RVSP, SBP and DBP of pulmonary artery, the right ventricular hypertrophy index, the thickening of arteriole in the heart and lung tissues were significantly higher in the MCT+SUC group than those in the MCT group(all P<0.05). HE staining showed slight thickening of the small arteries in the MCT group, the degree of thickening of the arterial wall of the MCT+SUC group was more obvious than that of the MCT group, and the infiltration of inflammatory cells in local myocardium increased. Sirius red staining and Masson staining showed abnormal proliferation of collagen fibers in the interstitial cells of the MCT+SUC group, and no abnormal collagen fibers in the MCT group. Western blot and immunohistochemistry showed that AngⅡ, FGF-2 and PDGF-BB expression levels in the MCT+SUC group and the MCT group were significantly higher than those in the control group, and the protein expressions of AngⅡ, FGF-2 and PDGF-BB in the MCT+SUC group increased more significantly than those in the MCT group. Conclusion: Succinic acid can exacerbate the formation of early fibrosis of cardiopulmonary arterioles in rats with pulmonary hypertension induced by monocrotaline. The mechanism may be due to regulating the expression of FGF-2 and PDGF-BB in the body, and eventually lead to increased expression of AngⅡ, causing interstitial fibrosis of cardio-pulmonary arteries.
Guo Yuanyuan , Fan Yuanyuan , Zhang Yongting , Wang Yu , Wang Yuanhui , Cai Jie , Li Jun , Zhang Lili
2017(9):1142-1147. DOI: 10.7655/NYDXBNS20170914
Abstract:Objective: To investigate whether the MSC-conditioned medium(MSC-CM) can protect injured liver and reduce liver fibrosis and its potential mechanisms. Methods: Six-week-old SD rats were allocated into three groups(each group n=8) as follows: model group; treatment group; normal group. The liver fibrosis model was established by intraperitoneal injection of low dose of CCL4(1.5 mL/kg)twice a week for eight weeks. MSCs were grown for 3~5 generation for the preparation of MSC-CM, which was concentrated 25-fold using ultrafiltration. From weeks 5 to 8, MSC-CM was injected every day with a dose of 2 mg/kg by tail vein in the treatment group; at the same time, the other two groups were injected with the same dose of L-DMEM. At the end of the experiment, hepatic stellate cells (HSCs) were isolated by in situ perfusion and density gradient centrifugation, and liver tissue was collected for pathologic analysis. The collagenous fiber was detected by Masson staining, while the expression of alpha-smooth muscle actin(α-SMA) in liver tissues was measured by immunohistochemical staining. To evaluate liver fibrosis, the gene and protein expression of α-SMA, transforming growth factor beta 1(TGF-β1), collagen I , matrix metalloproteinases-2(MMP-2), tissue inhibitor of metalloproteinases-2 (TIMP-2) in HSCs were evaluated by quantitative real-time PCR and Western blot. Results: In liver tissues, histological improvement was observed in hepatic fibrosis after MSC-CM treatment(P<0.01); the expression of α-SMA and collagenous fiber was significantly lower in the treatment group compared to the model group(P<0.05) , but not equal to the normal group. In HSC study, the mRNA expressions of TGF-β1,α-SMA,collagen Ⅰin the treatment group were significantly decreased than those in the model group(P<0.05); and their protein expressions were also lower in the treatment group(P<0.05),and the mRNA expression of MMP-2 increased compared to the model group. Conclusion: Our results showed that MSC-CM has a therapeutic effect on CCL4-induced liver fibrosis. And this process may be related to down-regulaing expression of TGF-β1, inhibiting HSCs activation and promoting collagen degradation.
Bu Hemei , Ma Xianghua , Sun Jin , Fang Da
2017(9):1148-1153. DOI: 10.7655/NYDXBNS20170915
Abstract:Objective: To detect the serum levels of retinol binding protein-4 (RBP-4) and glucose transporter-4 (GLUT-4) in the short-time (8 weeks) after duodenal-jejunal anastomosi (DJA) surgery and duodenal-jejunal bypass (DJB) surgery for type 2 diabetic rats,and attempt to explore theoretical basis and mechanisms of surgical treatment in type 2 diabetes. Methods: Type 2 diabetic rats were divided into the DJA surgery group (n=10), the DJB surgery group (n=10) and the sham surgery group (n=10). Weight, food intake, fasting blood glucose, fasting insulin, and RBP-4 level were measured before and after treatment. The glucose metablolism and insulin resistance before and after operation were evaluated. GLUT-4 level was detected at the 8th week after surgery. Results: Compared with the sham surgery group, the DJB and DJA groups showed decreased fasting blood glucose, reduced area under oral glucose tolerance test(OGTT) curve, improved insulin resistance, decreased BRP-4 level, and increased GLUT-4. Conclusion: DJA and DJB operation could significantly improve the glucose metabolism and insulin sensitivity of type 2 diabetic rats, which were independent of the weight loss, but related to decreased RBP-4 and increased GLUT-4 in skeletal muscle.
Yuan Hongjin , Lin Yuhui , Dong Jian , Zhu Dongya
2017(9):1154-1158. DOI: 10.7655/NYDXBNS20170916
Abstract:Objective:To explore the influence of intervention of histone deacetylase 2(HDAC2) on behavioristics of mice after photothrombotic stroke. Methods:Adeno-associated virus AAV-CAG-EGFP-Cre(2 μL) was microinjected through microcannula to the motor cortex of HDAC2flox/flox mice,aiming to knockdown HDAC2. And then,the expression of virus AAV-CAG-EGFP-Cre and its control AAV-CAG-EGFP was validated through immunofluorescence and Western blot. Seven days later,photothrombotic stroke was induced in the mice,and grid-walking task and cylinder task were conducted at day 3 before stroke and day 8 after stroke to detect behavior changes. Results:Compared with AAV-CAG-EGFP,AAV-CAG-EGFP-Cre significantly down-regulated HDAC2 expression,which was showed in the reduction of the number of GFP+/HDAC2+ cells and the expression level of HDAC2 in the peri-pinhole zone(P<0.05). Behavioral tests demonstrated that the decrease of HDAC2 didn't influence the behavior before ischemia,but significantly reduced foot faults in the grid-walking task(P<0.001) and asymmetry index in the cylinder task(P<0.001) after ischemia,which improved behavioral deficits. Conclusion:HDAC2 knock down by viral interference can promote functional recovery after photothrombotic stroke.
Xu Min , Han Xiaoning , Yang Zhongzhou , Zhao Chunjie , Gao Jun
2017(9):1159-1162. DOI: 10.7655/NYDXBNS20170917
Abstract:Objective: To investigate the function of 3-phosphoinositide-dependent protein kinase-1(PDK1) in the hippocampus dependent learning and memory. Methods: We used the Cre-Loxp to knock out PDK1 in empty spiracles homeobox 1(Emx1) positive cells conditionally, which is expressed in the dorsal telencephalon. PCR was used for genotyping and Morris maze was used to test the learning and memory of mice. Results: We found that the PDK1 knockout mice had weaker capacity of learning and memory associated with hippocampus. Conclusion: PDK1 plays an important role in the hippocampus-dependent learning and memory in mice.
2017(9):1163-1167. DOI: 10.7655/NYDXBNS20170918
Abstract:Objective: To establish an antral follicle culture system suitable for investigating oocyte maturation and ovulation in ICR mice, a strain of mice that are frequently used in biomedical research in China, and to study the effect of luteinizing hormone(LH) and epidermal growth factor(EGF) on induction of oocyte maturation and ovulation in this follicle culture system. Methods: Large antral follicles were dissected and cultured in vitro, oocyte maturation and ovulation in these cultured follicles were then induced by supplementation of LH and EGF to the culture medium. Results: The antral follicle culture system suitable for investigating oocyte maturation and ovulation in ICR mice was successfully established. In this culture system, treatment with LH or EGF alone was able to induce oocyte maturation and cumulus expansion in follicle-enclosed cumulus-oocyte complexes, whereas supplementation with both LH and EGF was necessary for induction of ovulation. Conclusion: LH/EGF induces oocyte maturation and ovulation in cultured large antral follicles of ICR mice.
2017(9):1168-1172. DOI: 10.7655/NYDXBNS20170919
Abstract:Objective:In order to improve the sensitivity and specificity of magnetic resonance contrast agent (MRCA) in the diagnosis of lesion site, and to overcome the limitations of single positive(T1) contrast agent and negative(T2) contrast agent as well as achieve the accurate diagnosis of diseases, a Fe3O4-BSAGd nanoparticle contrast agent was prepared for simultaneous T1-T2 dual-mode magnetic resonance imaging. The imaging performance and biocompatibility were evaluated. Methods:The superparamagnetic Fe3O4 nanoparticles with an oil-soluble particle size about 6 nm were prepared by the high temperature pyrolysis method, and the paramagnetic bovine serum albumin-gadolinium complex(BSAGd) was modified on the surface by phacoemulsification, while giving paramagnetic Fe3O4 nanoparticles, water solubility and good biocompatibility. Finally, the morphology, hydrated diametar(HD), saturation magnetization, relaxation efficiency(r), in vitro T1 and T2 weighted imaging and cytotoxicity were characterized. Results:The Fe3O4-BSAGd nanoparticles had a diameter of 10 nm and HD of about 32 nm, and the dispersion was uniform and stable. Fe3O4-BSAGd had a saturation magnetization of 30.2 emu/g. Longitudinal (r1) and transverse (r2) relaxation rates were 6.30 mM-1s-1 and 287.08 mM-1s-1, respectively. In vitro dual-mode imaging results showed a significant T1-T2 imaging effect and a concentration-dependent effect. The results of cytotoxicity test showed that the Fe3O4-BSAGd had a better biocompatibility. Conclusion:The effect of in vitro T1 and T2 weighted imaging of prepared Fe3O4-BSAGd nanoparticles was excellent with a good biocompatibility, which lays the foundation for the next step in vivo imaging.
Wu Tianyi , Bu Shoushan , Zhuang Hai , You Ying , Zhang Jisheng , Xu Yang
2017(9):1173-1176,1184. DOI: 10.7655/NYDXBNS20170920
Abstract:Objective: To study the preparation and properties of gelatin-graft-polyaniline(GP) hydrogel. Methods: GP was synthesized by direct doping technique, and camphor sulfonic acid(CSA) was used as dopant. The copolymer was dissolved in water and then cross-linked by EDC. The morphology and chemical feature, swelling ratios, dissolubility, biocompatibility and conductivity of GP hydrogels(GPH) were observed. Results: GPH had good conductivities (1×10-4~10-3 S/cm). The electrical property was enhanced with the content of polyaniline. Nonetheless, when the amount of polyaniline improved to a certain degree, the solubility was greatly reduced and swelling ratio was increased at the same time. Conclusion: GPH is a biodegradable conductive material and has great application prospects in tissue engineering.
Pu Chun , Qin Mingming , Liu Xia , Wang Wenjun , Gu Chunrong , Xu Jian , Wang Yue , Liu Jingping , Gu Min , Zhu Juanjuan , Pan Shiyang
2017(9):1177-1181. DOI: 10.7655/NYDXBNS20170921
Abstract:Objective: To investigate the expression of tumor specific protein 70 (SP70) in breast cancer tissues and blood to eva- luate whether SP70 could differentiate the benign breast tumor and malignant breast tumor. Methods: The SP70 expressions of paraffin sections in the tissues of 65 malignant breast tumors and 60 benign breast tumors were studied by immunohistochemistry(IHC) technique. The levels of SP70 in the serum of 58 malignant breast tumors, 62 benign breast tumors and 60 healthy subjects were studied by enzyme-linked immuno sorbent assay(ELISA). Results: The expression positive rate of SP70 was 55.38% in the tissues of malignant breast tumors, whereas, only 15.00% in benign breast tumors. The difference was statistically significant (χ2=23.846, P<0.001). The serum SP70 level of patients with malignant breast tumors was significantly higher than that in patients with benign breast tumors and healthy controls(F=11.884,P<0.001). The sensitivity of serum SP70 for diagnosis of malignant breast tumors was significantly higher than that of serum CA153 (53.45% vs. 44.82%), a significant association between serum SP70 and serum CA153 had not been found (r=0.132,P=0.324). The combine of serum SP70 and serum CA153 could significantly improve the sensitivity(84.38%). Conclusion: SP70 may be used to differentiate diagnosis of malignant breast tumors and benign breast diseases.
Cao Huan , Zhang Chengning , Sheng Yanhui , Kong Xiangqing , Sun Wei
2017(9):1188-1192. DOI: 10.7655/NYDXBNS20170924
Abstract:Objective: To identify risk factors associated with treatment resistant hypertension(TRH) and easily detectable through routine blood tests in chronic kidney disease(CKD) patients, who received hemodialysis. Methods: One hundred and seventy-four CKD patients with hypertension receiving hemodialysis between December 2009 and June 2015 were enrolled in this study. Of these patients, 69 patients had TRH; the other 105 patients had not. Univariate and multivariate logistic regression analysis was used to assess risk factors associated with TRH. Results: In our study population, the gender distribution, serum level of creatinine and hemodialysis duration were comparable between two groups. The mean age,serum leves of uric acid(UA), white blood cell numbers, aspertate aminotransferase(AST),gamma-glutamyl transferase(GGT) and adenosine deaminase(ADA) were lower in the case group than those in the control group. The serum level of calcium and albumin(ALB) was higher in the case group than that in the control group. Age younger than 40(OR:3.350,P=0.010),GGT less than 20 U/L(OR:2.554,P=0.024), 10≤AST<15 U/L(OR:2.499,P=0.011), and ADA less than 12 U/L(OR:2.259,P=0.044) were positively correlated with TRH. Meanwhile, serum level of ALB less than 41 g/L(OR:0.298,P=0.002) was negatively correlated with TRH. Stepwise multi-variate logistic regression showed that ALB was positively(OR:1.130,P=0.002) and ADA was negatively(OR:0.900,P=0.027) correlated with TRH. Conclusion: The younger age(<40 years) and lower serum level of GGT(<20 U/L), AST(10 U/L≤AST<15 U/L) and ADA(<12 U/L) are risk factors, and serum levels of ADA decreasing and ALB increasing are independent risk factors for TRH in hemodialysis patients.
Zhang Yu , Zhang Hongwen , Yang Jin , Wu Yanqing , Tan Ruoyun , Wang Libin , Liu Yun , Wang Yuanyuan , Wei Jifu , Wang Yongqing
2017(9):1193-1199. DOI: 10.7655/NYDXBNS20170925
Abstract:Objective: To construct a population pharmacokinetics(PPK) model of sirolimus in Chinese renal transplantation patients, study the pharmacological characteristics of sirolimus, and provide theory support for personalized drug use. Methods: One hundred and eleven renal transplantation patients, who were orally administrated with sirolimus after transplantation, were enrolled in the study. Steady-state concentrations of sirolimus and related laboratory test results were retrospectively collected. Phoenix NLME was used to perform a population pharmacokinetics model. Influence factors on pharmacokinetic parameter were analyzed. Performance of final model was internally assessed by bootstrapping and visual predictive checking(VPC). Results:A one-compartment model with first-order elimination pharmacokinetics provided the best fitting. In all fixed effects, hematocrit influenced the clearance of sirolimus, alkaline phosphatase influenced volume of distribution. Typical value of CL/F was 10.8 L/h, typical value of V/F was 1011 L. Stability and predictive performance were accepted by bootstrapping and VPC. Conclusion: A PPK model of sirolimus in Chinese renal transplantation patients is successfully established in this study. Effects of weight, age, gender, drug dose, drug combination, liver and kidney function on pharmacokinetic parameter were inspected. It is of great significance for clinical rational drug use of sirolimus.
Gu Lihua , Huang Xiaodan , Ji Guozhong
2017(9):1222-1226. DOI: 10.7655/NYDXBNS20170932
Abstract:Objective:To assess the association between miRNA-146a rs2910164 polymorphism and HCC risk by a meta-analysis. Methods:A systematical search of studies was conducted in papers published in PubMed , Embase ,Web of Science, China National Knowledge Infrastructure(CNKI) and Chinese Biomedical Literature Database(CBM) databases before January 30, 2017. Odds ratios(OR) and 95% confidence intervals(95% CI) were used to evaluate the association between miRNA-146a polymorphism and HCC risk. Results:A total of 18 case-control studies involving 5 657 cases and 6 680 healthy controls were included in this meta-analysis, in which no association was found between miRNA-146a rs2910164 polymorphism and HCC risk in any genetic models(C versus G:OR=0.96, 95%CI=0.88~1.06;GC versus GG:OR=0.98,95%CI=0.88~1.09;CC versus GG:OR=0.90,95%CI=0.74~1.09;GC/CC versus GG:OR=0.95,95%CI=0.82~1.10;CC versus GC/GG:OR=0.95,95%CI=0.84~1.07). Conclusion: No sufficient evidence supports the association between miRNA-146a rs2910164 polymorphism with hepatocellular cacinoma risk. The conclusion should be verified in large-scale and well-designed studies in the future.