Table of Content

05 January 2016, Volume 36 Issue 1

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Recombination Onconase promotes the apoptosis of rat hepatoma cell RH-35

2016, 36(1):  1-5. 

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Objective To investigate the apoptosis and cell growth inhibition in vitro effects of recombination onconase (rONC) on rat hepatoma cell RH-35. Methods After RH-35 cell in vitro were treated with various concentrations of rONC. Cel1 proliferation were determined by the SRB method and the bioactivities of these cell were detected by Hoechst33258. The effect of rONC on cell cycle were detected by flow cytometry assay. The expression levels of cell proliferation related genes and proteins after RH-35 cell treated with rONC were detected by qRT-PCR and Western blot. Results The proliferation of RH-35 cell were significantly inhibited after treated with rONC for 24 hours and this inhibitory action was positive correlated with time and dose-dependent (P<0.05) . The value of IC50 was 7.38 ?mol/L. The nucleus appeared some characteristic of apoptosis after RH-35 cell treated with rONC .The proportion of cells in G0/G1 phases remarkably rise after 24 h treatment with rONC, rONC could down-regulate the expression level of PCNA, CCNA2, BCL-2 and up-regulate BAX gene levels and protein levels. Conclusion rONC could inhibit the growth and induce apoptosis of RH-35 cell in vitro due to the down-regulated expression of PCNA, CCNA2, BCL-2 and up-regulated expression of BAX.

The expression of c-Myc protein in acute promyelocytic leukemia cell lines is inhibited by recombinant lentivirus expressing PML(?NLS)

2016, 36(1):  6-11. 

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Objective To construct recombinant lentivirus carrying PML(?NLS) gene and observe its effect on the expression of c-Myc protein in leukemia NB4 cell lines. Methods PML(?NLS) gene was amplified by PCR using plasmid pCMV-HA-PML(?NLS) as a template and cloned into the lentivirus vector LV5-EF1a-GFP/puro.The recombinant lentiviral vector，pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells，packaging virus. Then NB4 cells were infected with the supernatant containing lentiviral particles，and its infection efficiency was determined by fluorescent microscope，PML(?NLS) mRNA transcription and protein expression in NB4 cells were determined by RT-PCR and Western blot respectively，the expression of ?-catenin，γ-catenin and c-Myc proteins in NB4 cells were analyzed by Western blot. Cell proliferation was detected by CCK-8. Results Recombinant lentivirus LV5-PML(?NLS) was successfully constructed. The infection efficiency in NB4 cells reached 82.74％，the PML(?NLS) gene in LV5-PML(?NLS) was expressed stably in NB4 cells； the expression of β-catenin, γ-catenin and c-Myc proteins were decreased in NB4 cells infected with LV5-PML(?NLS). The activity of cell proliferation in NB4 cells was significantly enhanced as compared to the untransfeced and empty viral groups. Conclusion Recombinant lentivirus LV5-PML(?NLS) was constructed successfully and efficiency infected in NB4 cells. In addition，it could inhibit the expression of c-Myc protein.

Aging model bone marrow stromal cells inhibits proliferation and differentiation of hematopoietic cells

2016, 36(1):  12-18. 

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Objective: To explore the influence of senescence-bone marrow stromal cells on the proliferation and differentiation of hematopoietic cells and to provide the theoretic and experimental evidences for explaining the effect of senescent hematopoietic microenvironment on Hematopoietic stem/progenitor cells. Methods: The bone marrow stromal cells (BMSCs) were isolated by whole bone marrow adherent culture from rats, the experiment cells were divided into control group and aging group. The control group was cultured as usual and the aging group was cultured with D-galactose (D-Gal) 30mg/mL for 48 hours. And then the proliferation ability of BMSCs was detected by Cell Counting Kit-8(CCK-8); the distribution of cell cycle was analyzed by flow cytometry (FCM); the senescence-associated β-galactosidase（SA-β-Gal）staining was used to detect the senescent BMSCs; Western Blotting examined the expression level of P16, P21and P53. The bone marrow hematopoietic cells were co-cultured with BMSCs, the proliferation and differentiation ability of CFU-Mix was detected by colony forming assay. The amounts of SCF、GM-CSF and IL-1β in BMSCs culture supernatant were detected by ELISA; DCFH-DA fluorescent staining and FCM analyzed the level of ROS (reactive oxygen species) in BMSCs; MDA (malonaldehyde) content and total SOD (superoxide dismutase) activity was analyzed as well using enzymatic assay. Results: Compared with the control group, D-Gal induced senescent BMSCs displayed a decrease in proliferation; the BMSCs were hold in G0/G1 phase arrest(p<0.01); the positive ratio of SA-β-Gal stained BMSCs also significantly increased; The expression of senescence-related proteins including P16、P21 and P53 were obviously up-regulated(p<0.01). The proliferation and differentiation ability of the hematopoietic cells co-cultured with aging BMSCs declined. In the aging BMSCs, ROS and MDA the index of oxidative damage increased and SOD the antioxidant index declined(p<0.01). The amount of IL-1β、GM-CSF、SCF in BMSCs culture supernatant of aging group decreased(p<0.01) .Conclusion: Senescent BMSCs inhibit proliferation and differentiation of the hematopoietic cells and the underling mechanism may be related to the oxidative damage of BMSCs, thus reduced secretion of cytokines by BMSCs.

IL-2 inhibits the differentiation of mouse CD4+CD62L+T cells into Th17

2016, 36(1):  19-23. 

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Objective To investigate the effect of IL-2 on the differentiation of mice spleen CD4+CD62L+T cells into Th17 in vitro. Methods CD4+CD62L+T cells from C57BL/6 mice with MACS were divided into two groups, control group and IL-2 treated group, to be cultured in anti-CD3 and anti-CD28 antibody coated plate for 3 days. Control group cells were cultured in classical Th17 conditional medium, while IL-2 was added into the conditional medium of IL-2 treated group. Cell proliferation, cell apoptosis, the concentration of cytokine IL-17A, expression level of specific transcription fator Rorγt, and percentage of CD4+IL-17+Th17 were measured by CFSE fluorescent staining, Annexin V-PI, ELISA, qRT-PCR, and FACS, respectively. Results The purity of separated CD4+CD62L+na?veT was more than 95%. Compared with control group, the proliferation was improved (P<0.05) with a reducing apoptosis (P<0.05) in IL-2 treated cells. Moreover, IL-2 treated cells produced significantly lower concentration of IL-17A (P<0.05), decreased expression level of Rorγt (P<0.05), and less CD4+IL17+cells (P<0.05) compared with control group. Conclusion During the process of CD4+CD62L+T differentiating into Th17 in vitro, IL-2 could facilitate T cell proliferation but inhibit the differentiation of Th17.

Prostaglandin D2 stimulates human airway mucus hypersecretion mediated by DP2

2016, 36(1):  24-29. 

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Objective To investigate the effect of D prostanoid receptor 2 （DP2） on mucin (MUC) 5AC overexpression induced by prostaglandin D2（PGD2）and the potential mechanism． Methods The human airway epithelial（16HBE）cells were stimulated with PGD2 (1 μmol/L). In intervention experiments, cells were pretreated with DP2 antagonist AZD6430, DP1 antagonist BWA868C and PI3K specific inhibitor LY294002. Then, PGD2 stimulation was used. Cells were divided into 6 groups: control group ( incubated in DEME / Ham's F12 medium without fetal bovine serum), PGD2 stimulation group, PGD2 stimulation+AZD6430 group, PGD2 stimulation+BWA868C group, PGD2 stimulation +LY294002 group. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)- 4 and IL-13 protein were tested by real-time PCR and ELISA, respectively. MUC5AC protein level was assessed by immunofluorescence. Western blot was used to detected the levels of DP2, DP1, PI3K and phosphorylation of AKT (p-AKT). MUC5AC mRNA expression was tested by real-time PCR. Results PGD2 caused a obversely increase of MUC5AC, TNF-α, IL-4, IL-13, DP2, PI3K and p-AKT when compared with normal control group (P<0.05). Inhibited of DP2 activity, production of TNF-α, IL-4, IL-13, MUC5AC, PI3K and p-AKT were significantly lower than that of PGD2 stimulation group (P<0.05). The level of TNF-α，IL-4, IL-13 and MUC5AC mRAN and protein expression were also attenuated after treatment with LY294002 (P<0.05). Conclusion Our study suggests that DP2 mediates mucus hypersecretion induced by PGD2 via PI3K/AKT pathway.

CGRP inhibits the hypoxia-induced apoptosis via NOS/NO pathway in cardiomyocytes of rat

2016, 36(1):  30-34. 

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Objective To investigate whether Calcitonin Gene-Related Peptide (CGRP) plays a protective role in cardiomyocytes against hypoxia-induced apoptosis via an NO-mediated pathway. Methods H9c2 cardiac cells were exposed to hypoxia for 2 h to establish a model of myocardial hypoxic-ischemic injury. The cells were pretreated with either CGRP or an NOS inhibitor (L-NAME) before being exposed to hypoxia for 30 min. Cell viability was analyzed using a cell counter kit 8 (CCK-8). The expression levels of several apoptosis proteins (p53, caspase-3, cytochrome C) and nitric oxide synthase (NOS) were detected via a Western blot assay. An NO kit was used to evaluate the production of NO. Results Our studies demonstrated that pretreatment of H9c2 cardiac cells with CGRP for 30 min prior to exposure to hypoxia markedly improved cell viability; the same effect was observed following pretreatment with the NOS inhibitor, L-NAME. Additionally, CGRP enhanced the expression of both eNOS and p-eNOS; the application of both L-NAME and CGRP attenuated the hypoxia-induced expression of iNOS and enhanced the hypoxia-mediated decrease in NO. Furthermore, CGRP significantly decreased the hypoxia-induced overexpression of several apoptotic proteins, p53, caspase-3 and cyto C. Interestingly, the expression levels of cell death, NOS and apoptotic factors were slightly attenuated in the presence of L-NAME and CGRP co-working 2 hours of acute hypoxia. Conclusion CGRP protects cardiomyocytes from hypoxia-induced apoptosis via the modulation of the production of NO.

Construction of Recombinant Lentiviral Vector and Neutrophil Elastase Promotes Proliferation in NB4 Cell

2016, 36(1):  35-40. 

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Objective To construct recombinant lentiviral LV5-NE and screen NB4/LV5-NE cell line. Explore the effects of neutrophil elastase on proliferation and apoptosis in NB4 cell. Method Calculate the title of LV5-NE by fluorescence counting. There were three groups (blank control group, LV5-NC group, LV5-NE), the expression level of Western blot was used to detect the expression level of NE protein and its cleaved products in NB4 cell after infected by LV5-NE. CCK-8 was used for cell proliferation analysis. Flow cytometry was used for cell cycle and apoptosis analysis. Western blot was used to detect the expression level of phosphorylated Akt, Akt ，Bax and Bcl2 in NB4 cell. Results It，s success to construct lentivir of LV5-NE and screen NB4/LV5-NE cells. Neutrophil elastase obviously promotes proliferation and inhibits apoptosis in NB4 cell（P<0.05）. Conclusion Neutrophil elastase may active PI3K/Akt pathway to promote proliferation and inhibit apoptosis in NB4 cell.

NLS-RARα regulates proliferation of leukemia cell line NB4 through the activation of AKT

2016, 36(1):  41-46. 

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Objective To construct NLS-RARα recombinant lentivirus and detect its effect on proliferation and AKT protein of leukemia cell line NB4. Methods NLS-RARα gene was amplified by PCR using plasmid pCMV-HA-NLS-RARα as a template. The resulting gene of NLS-RARα was subcloned into the shuttle plasmid LV5-EF1a-GFP/PURO by using DNA recombinant technique. The four plasmids system was transfected into 293T cells by liposome. The virus supernatant was harvested and concentrated, then determined the titer. The recombinant lentivirus vector of NLS-RARα was used to infect Leukemia cell line NB4, the stably transfected cell lines were selected by puromycin. Cell viability was detected by CCK-8 assay. Cell cycle and infection efficiency were measured by flow cytometry. The expression of NLS-RARα, t-AKT and p-AKT were detected by Western blot. Results Recombinant lentivirus plasmid LV5-NLS-RARα was constructed correctly as proved by DNA sequencing. The titer of the lentivirus reached 2×108 Tu/mL. The infection efficiency of LV5-NLS-RARα in NB4 cells reached 86.54%. The proliferation of NB4 cells which over-expressed NLS-RARα was enhanced. The proportion of cells in S phase was increased which was decreased in G1 and G2 phases of the cell cycle. The protein level of NLS-RARα and p-AKT were significantly increased. The level of p-AKT was decreased after pretreatment with PI3K inhibitor LY294002. Conclusion NLS-RARα prometos proliferation of leukemia cell line NB4 through the activation of AKT.

Effects of Salidroside on cell proliferation and cycle in mouse mesenchymal stem cells

2016, 36(1):  47-52. 

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Objective To investigate the effect and mechanism of salidroside on cell cycle and proliferation via ERK signal pathway in mouse marrow mesenchymal stem cells. Methods The experiment including control group (D-MEM/F-12 complete medium), different concentration of salidroside induced group (5~100 μg/mL) and blocked group (30 μg/L PD98059 + 5~100 μg/mL). The cell cycle, inhibition rates and the expression levels of cell cycle-related proteins were detected by flow cytometry, CCK8 method and western blot respectively. Results The cell proliferation were inhited in 10 μg/mL induced group and blocked group, meanwhile the inhibition rates in 5 μg/mL (P<0.01), 25 μg/mL (P<0.05) and 100 μg/mL (P<0.05) induced groups were significantly higher than those in blocked groups. The G0/G1 phase cell rate in 25 μg/mL induced group and blocked group were both increased. The expression of CyclinA and CyclinD1 protein were downregulated, whereas p21 protein expression was upregulated in both induced group and blocked group to compared with the control group. The expression of CyclinA protein was downregulated in blocked group to compared with the induced group, the expression of CyclinD1 protein was downregulated in 50 and 100 μg/mL induced groups to compared with blocked group, whereas the expression of p21 protein in induced group was extremely higher than blocked group. Conclusions Salidroside could inhibit MSCs proliferation and affect cell cycle via regulating the expression of cell cycle-related proteins.

The expression of aquaporin 1 and 5 in the LPS-induced lung injury in rats

2016, 36(1):  53-57. 

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Objective To investigate the expression and significance of the Aquaporin (AQP) 1, 5 in the LPS-induced lung injury in rats. Methods 48 Wistar rats were divided into control-group, ALI-group. Preparation with acute lung injury (ALI) model by intravenous injection of LPS in rats, control-group injected saline. Rats were sacrificed in 2, 6, 12 and 24h, and the samples were collected after the successful modeling. Observing lung HE staining morphological, determining lung wet/dry weight ratio (W/D) and lung permeability index, ELISA detecting serum TNF alpha, MIP-1 alpha level. At the same time, analysis AQP1, AQP5 protein and mRNA content changes through Western-blot, Immunohistochemistry and Real-Time PCR method. Results Compared with control-group, the TNF alpha, MIP-1 alpha level in ALI-group were significantly elevated at 2, 6 and 12h(P<0.05), and to 24h point gradually down to normal level. HE staining showed that the alveolar and interstitial edema and inflammatory cell infiltration after LPS injection 2h, most obviously in 12h. The lung W/D, BALF protein and pulmonary permeability index in ALI-group were significantly higher than those of the control-group (P<0.05), and reached the peak in 12h. At the same time, the expression levels of AQP1, AQP5 in ALI-group were significantly lower than that in control-group (P<0.01). The correlation analysis showed that the expression of AQP1 and AQP5 mRNA was negatively correlated with the lung W/D ratio, serum TNF alpha and MIP-1 alpha level. Conclusion LPS can induces rats acute lung injury, The formation of pulmonary edema may be related to the down-regulation of AQP1 and AQP5.

Grape seed procyanidin alleviates renal function and structure injury in SHR

2016, 36(1):  58-61. 

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Objective To observe the effect of grape seed procyanidins (GSP) on renal function and structure in spontaneous hypertensive rats (SHR) and to discuss the possible mechanism. Methods 24 male SHR (8-week-old) were randomized into 4 groups (n=6 each group): SHR , low and high dose GSP group (50 and 200 mg/kg) , captopril used for positive control group (30 mg/kg), meanwhile, 6 Wistar Kyoto rats (8-week-old) were randomly served as control group. After 6 weeks treatment, tail systolic pressure, urea nitrogen (BUN) and creatinine (SCr) contents in serum, malondiadehyde (MDA) content, superoxide dismutase (SOD) and catalase (CAT) activity in the renal cortex of the rats were measured respectively. Cystain C (Cys C) level in serum was detected by ELISA. Changes of renal structure were observed after HE staining. Western blot was performed to detect the expressions of ERK1/2 protein in the renal cortex. Results GSP could significantly decrease tail systolic pressure (P<0.01), improve renal function parameters, reduce MDA content and the expression of ERK1/2 in renal cortex (P<0.05), but increase SOD and CAT activity (P<0.05), relieve renal structure injury, especially in high dose GSP group (P<0.01). Conclusion GSP has a beneficial effect on renal function and structure injury in SHR, and its mechanism may be involved in GSP could lower the SBP, inhibit oxidative stress and reduce expression of ERK1/2 in renal cortex.

Antitumor effect of human amniotic epithelial cells supernatant on human hepatoma BEL-7402 cells in vitro

2016, 36(1):  62-67. 

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Objective To investigate the migration, proliferation and cell apoptosis effects of human amniotic epithelial cells(hAECs) supernatant on human hepatoma BEL-7402 cells cultured in vitro. Methods BEL-7402 cells were randomly divided into five groups.The control group contained no hAECs supernatant, while the other four treatment groups were added different volume fraction of hAECs supernatant namely 6.25% ,12.5% ,25% and 50% for 24 hours.The scratch assay,MTT assay and flow cytometry were used to detect the migration, proliferation and apoptosis of BEL-7402 cells. The expression levels of apoptosis-related proteins were detected by Western blot method. Results hAECs supernatant could dose-dependently inhibit the migration and proliferation of BEL-7402 cells and induce apoptosis (P<0.05).The expressions of cleaved caspase-3 and cleaved caspase-8 of BEL-7402 cells in 25% and 50% hAECs supernatant groups were higher than those in control group(P<0.05). Conclusion hAECs supernatant has some antitumor effects on BEL -7402 cells cultured in vitro.

HBSP alleviates the rat myocardial cell injury induced by Anoxia/Reoxygenation

2016, 36(1):  68-72. 

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Objective: To study the effection of Helix B surface peptide(HBSP) to myocardial apoptosis in the state of acute anoxia / reoxygenation and its possible mechanism. METHODS: CμLtured neonatal rat cardiomyocytes H9C2 cell line in vitro and establishment anoxia (3h) / reoxygenation (3h) model. The experiment were divided into three group: control group, anoxia / reoxygenation group (A/R) and H / R + HBSP groups. CμLture supernatants were collected to measure the lactate dehydrogenase (LDH) levels and the myocardial apoptosis rate was detected by using Annexin-V and PI double staining and flow cytometry. What’s more, Western blot analysis was used to measure the expression of cytochrome C, and the activities of caspase-3 and caspase-9 were determined by a colorimetric method. ResμLts: Compared with the control group, cell viability and mitochondrial cytochrome C protein levels in A/R group were significantly decreased（P<0.01）, while the rate of apoptosis, caspase-9, caspase-3 activity and cytosolic cytochrome C protein levels were significantly increased（P <0.01）. Conclusion: HBSP significantly protect neonatal rat cardiomyocytes from the injury of anoxia /reoxygenation and the mechanism may be inhibiting the cell apoptotic mediated by mitochondrial pathway.

Effects of down-regulated long non-coding RNA PVT1 expression on cell apoptosis, migration and invasion of pancreatic cancer cells BxPC-3

2016, 36(1):  73-79. 

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Objective 　To study the effects of down-regulated long non-coding RNA PVT1 expression on proliferation, apoptosis, migration and invasion of pancreatic cancer cells. Methods PVT1 interference sequences siPVT1-1 and siPVT1-2 were transfected into pancreatic cancer cell line BxPC-3 in vitro, and the siPVT1-NC was taken as negative control. The ability of proliferation and apoptosis of BxPC-3 were detected by CCK-8 assay and Flow cytometry, respectively. Wound healing assay and Transwell assay were study the ability of migration and invasion. The mRNA expression of E-cadherin, N-cadherin, β-catenin, vimentin and PVT1 were detected by qRT-PCR. Apoptosis related protein (Bax, Bcl-2, Caspase 3) and epithelial-mesenchymal transition related protein (E-cadherin, N-cadherin, β-catenin, vimentin) were detected by Western blot. Result Down-regulated expression of pancreatic cancer cell line BxPC-3 reduced the ability of proliferation and promoted cell apoptosis (P<0.05). The number of migrated and invased cells were reduced significantly when down-regulated the expression of PVT1 (P<0.05). After transfected with siPVT1-1 and siPVT1-2, the molecular and protein level of E-cadherin were increased, while N-cadherin, β-catenin and vimentin were reduced (P<0.05). The protein level of Bax and Caspase 3 were increased, Bcl-2 was reduced (P<0.05). Conclusion Down-regulated expression of long non-coding RNA PVT1 can reduce cell proliferation and promote cell apoptosis, meanwhile, the ability of migration and invasion are also inhibited. Down-regulated expression of PVT1 can reverse epithelial-mesenchymal transition of pancreatic cancer cells.

Vitamin E has antioxidant effect both in vivo and in vitro

2016, 36(1):  80-84. 

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Objective:To explore the antioxidant effect of Vitamin E on both in the rat in vivo and in hepatic RBL cells in vitro.Methods:The cultured human hepatic RBL cells were exposed to H2O2 as an oxidant and treated with 0.4mM doze of Vitamin E before or after exposure of H2O2. The cell survival and apoptosis were detected by MTT and TUNEL assays. The expression level of NF-kB，Bcl-2，Bax，Hsp-70 and Caspase-3 was determined by immunohistochmistry and Western blotting. Will clean level only 20 male Wistar rats were divided into control group and the big and small doses of vitamins E35 and 15 mg/kg 2 ml lavage solution，1 times a day，a total of three times，the biochemical method to detect within the plasma after 3 and 6 d T – AOC，SOD，GSH and MDA levels. Results: H2O2 damage group (Ec) apoptosis rate increased (P< 0.01)，intracellular Bax，HSP – 70，the nf-kappa B and caspase 3 significantly higher (P< 0.01)，while the Bcl - 2 decreased significantly (P< 0.01)，vitamin E after the intervention can significantly alleviate the above changes (P< 0.01)，H2O2 damage before intervention is better than that after intervention. Lavage after 3 d plasma T - the AOC，SOD，higher level of GSH，MDA lowered (P< 0.01)，and 6 d after the relevant index change is more significant (P< 0.05). Conclusion: The results showed that the Vitamin E displays anti-oxidant capacity through upregulating the antioxidant enzyme system and regulating the expression of relevant proteins.

Down-regulation of RNF167 suppresses the production of IFN-β in viral-infected mouse peritoneal macrophages

2016, 36(1):  85-88. 

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Objective To investigate the regulation of IFN-β expression by RNF167 in viral-infeted macrophages. Methods The expression of RNF167 was detected by Western blot in mouse peritoneal macrophages infected with either RNA virus VSV or DNA virus HSV-1. The expression of IFN-β was determined by real-time quantitive PCR (q-PCR) and ELISA assay in macrophage transfected with si-RNF167 and viral infection. P-IRF3 level was analyzed by Western blot. Results RNF167 was down-regulated in VSV infected mouse peritoneal macrophages （P < 0.01）. Knockdown of RNF167 by si-RNF167 significantly decreased the expression of IFN-β and the level of p-IRF3 in RNA viral-infected cells （p < 0.01, but not in HSV-1-infected cells. Conclusion With RNA viral infection, the level of RNF167 expression was decreased, which specifically regulated IFN-β expression in mouse peritoneal macrophages.

SIRT1 overexpression inhibits tunicamycin-induced endoplasmic reticulum stress in chondrocytes

2016, 36(1):  89-93. 

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Objective To explore the effect of silent mating type information regulation 2 homolog 1 (SIRT1) on tunicamycin-induced endoplasmic reticulum stress (ERS) in chondrocytes. Methods After isolation of human normal chondrocytes and osteoarthritis (OA) chondrocytes, morphological identification was performed, collagen type II expression was determined by immunohistochemical staining and SIRT1 expression was detected by Western blot. Cultured chondrocytes were divided into five groups: control group (OA chondrocytes, normal chondrocytes), normal chondrocytes group (Tuni, SIRT1 overexpression, and Tuni + SIRT1 overexpression). After 24 h treatment, the expression of ERS-related proteins of C/EBP homologous protein (CHOP), eukaryotic initiation factor-2 α (eIF2α) phosphorylation, and activating transcription factor 4 (ATF4), and the expression of apoptosis-related proteins of Bcl-2, Bax, and Caspase-3, as well as p-p38 protein were determined by Western blot. NF-κB activity was determined by ELISA. Results Compared to human normal chondrocytes, both the cell proliferation and the expression of collagen type II and SIRT1 in OA chondrocytes were markedly reduced (P<0.05). OA control group and 0.5 mg/L tunicamycin treatment consistently resulted in ERS and cell apoptosis with concomitant enhancement of CHOP, p-eIF2α and ATF4 proteins, increases of NF-κB activity and p-p38, Bax, Caspase-3 proteins, and reduction of Bcl-2 (P<0.05). However, SIRT1 overexpression could reverse these effects. Conclusion SIRT1 overexpression inhibits tunicamycin-induced ERS and cell apoptosis, and further attenuates p38/NF-κB activation in chondrocytes.

The expression and effect of long non-coding RNA HOTAIR in cervical cancer patients and HeLa cells

2016, 36(1):  94-98. 

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Objective To investigate the expression of long non-coding RNA HOTAIR in cervical cancer tissues and its effects on HeLa cell growth and apoptosis. Methods The expression of HOTAIR in 66 cervical cancer tissues and matched adjacent normal tissues were measured using quantitative real-time RT-PCR and its correlation with the clinical parameters was analyzed. Lentiviral mediated shRNA was used to knockdown HOTAIR in HeLa cells to investigate its role in cell growth and apoptosis. Results HOTAIR expression in cervical cancer tissues was significantly upregulated compared with the matched nontumorous tissues (8.25±0.46 vs. 3.6±0.26; P<0.001); Increased HOTAIR expression was significantly correlated with tumor size(P<0.05), tumor stage(P<0.05) and lymph node metastasis(P<0.05). Knockdown of HOTAIR significantly reduce cell growth, cell cycle arrest at G0/G1 phase, and promoted cell apoptosis. Conclusion High expression of HOTAIR is involved in cervical cancer progression and could be a potential target for diagnosis and gene therapy.

Expression of ANKRD49 in non-small cell lung cancer and its clinical significance

2016, 36(1):  99-103. 

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Objective To investigate the expression levels of ANKRD49 in non-small cell lung cancer (NSCLS) and its clinical significance. Methods The specimens of carcinoma tissues and their paired para-carcinoma tissues were collected from 75 NSCLS patients and were made into tissue microarray. Immunohistochemistry was applied to detect the expression level of ANKRD49 in these specimens. The correlation between the expression level of ANKRD49 and the different clinical parameters of NSCLC was analyzed. Western blot assay was used to detect the expression level of ANKRD49 in human airway epithelial cell line (HBEC) and different lung cancer cell lines. Results The positive expression rate of ANKRD49 in the carcinoma tissues （82.67%）was significantly higher than that in the para-carcinoma tissues （49.33%）（P ＜ 0. 05）, and the positive expression rate of ANKRD49 in poorly differentiated carcinoma tissues （80%）was significantly higher than that in well differentiated carcinoma tissues （42.6%）（P ＜ 0. 05）. ANKRD49 was expressed only in A549 and NCI-H1650 lung adenocarcinoma cell lines rather than in normal airway epithelial cell line and other lung cancer cell lines. Conclusion ANKRD49 may be involved in the progression of NSCLC.

Disorders of Sexual Development caused by haploinsufficiency of DMRT1 gene

2016, 36(1):  104-106. 

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Objective: To increase the knowledge on disorders of sex development (DSD) caused by DMRT1 haploinsufficiency. Methods: Clinical features in a patient with 46XY DSD were described. Peripheral lymphocytic karyotype was measured in this patient and her parents. Furthermore, array comparative genomic hybridization (aCGH) was done to the patient. Results: (1) Clinical features: the patient presented with female genital, and neither uterus nor ovaries were detected by B ultrasound. Mental and physical development retardation was observed, consisted with the manifestation of 9p deletion syndrome. (2) Karyotype: Her mother was 46XX,t(7;9)(q35,p24); Her father was 46XY; The patient was 46XY, der(9) t(7;9) (q35,p24); (3) aCGH scan: 14.37Mb triplication was fond in the terminal of 7q (144741153-159098761); 9.72Mb deletion was found in the terminal of 9p (10001-9733061), containing the following genes, EZH2, MNX1, DMRT1, DMRT2, SHH, SMARCA2, GLDC, VLDLR, DOCK8 and GLIS3; Conclusions: DMRT1 gene plays an essential role in gonadal development. Haploinsufficiency in this gene may result in testicular dysplasia and DSD.

Sinomenine down-regulates the expression of adipokines receptor in synovium of rabbit knee osteoarthritis model

2016, 36(1):  107-108. 

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Inhibitory Effect of Diallyl trisulfide on humam multiple myeloma H929 cells proliferation

2016, 36(1):  109-111. 

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The role of AXL in tumor metastasis and drug resistance and strategy in anti-tumor therapies

2016, 36(1):  112-115. 

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The receptor tyrosine kinase (RTK) AXL is over expressed in multiple tumor cells and mediates the metastasis and drug resistance of tumor cells through a variety of signaling pathways, especially form positive feedback loops with EMT, therefore has become a new tumor therapeutic target. By inhibiting the expression of AXL can effectively reverse EMT and tumor metastasis and drug-resistance.

A novel mechanism of TFEB regulating lipid metabolism for stabilizing atherosclerotic plagues

2016, 36(1):  116-120. 

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TFEB could induced by autophagic regulatory factors, such as mTOR and cellular stresses. TFEB could media the translation of peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PPAR α), peroxisome proliferator-activated receptor γ coactivator 1 α(PGC-1α) and autophagy, lysosomal synthesis related genes, then regulat lipid metabolism and the release inflammatory cytokine. AS a novel Mechanism, TFEB could regulate Lipid metabolism in order to restrain the progression of atherosclerosis and promote to plaque stabilization.

Research progress on hypercoagulable state in Cushing’s syndrome

2016, 36(1):  121-124. 

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Cushing’s syndrome (CS) results from chronic exposure to excessive levels of glucocorticoids. Generally it is accompanied by a hypercoagulable state. Patients with CS have a higher risk of arterial and venous thrombotic events, cardiovascular disease morbidity and mortality. This review aims to introduce its pathophysiological mechanisms, and to provide examples and guidelines for antithrombotic prophylaxis in CS.

Correlation research on immunosenescence and cells of the innate immune system

2016, 36(1):  125-129. 

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Immunosenescence is one of the most important biological changes occurring during human aging . It affects both innate and adaptive immunity . The impact of aging on innate immunity is less well understood particularly in humans. There are modifications in the number, surface molecules ,cytokines and signal transduction as well as the function of cells of the innate immune system during the ageing process.

The impact of integration of early scientific research training on undergraduate diagnostic medical education

2016, 36(1):  130-133. 

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Objective: To evaluate the impact of integration of early scientific research training on undergraduate diagnostic medical education. Methods: Undergraduate medical students from Tsinghua University who received 2-year early scientific research training were arranged to received diagnostic medical classes together with the medical students from Peking Union Medical College (PUMC) who received similar preclinical courses except no scientific training. The scores of clinical diagnostic knowledge, clinical laboratory diagnostics, history taking and physical examination were compared between the two groups. Results: The mean age of the 16 students received early scientific research training was 23.5±0.5 years old．There were 70 students from the PUMC group without scientific training. Their mean age was 22.4±0.7 years old. The results showed that mean scores of clinical diagnostic knowledge were significantly higher in the group received early scientific research training（p<0.05）. And no group differences were found for the scores of clinical laboratory diagnostics（p=0.239）, history taking and physical examination. Conclusions: Early scientific research training may benefit diagnostic knowledge acquisition.

A survey on understanding of ultrasound examination among resident physicians in PUMC Hospital

2016, 36(1):  134-137. 

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Objective To describe the knowledge of ultrasound examination for resident physicians. Methods A? cross-sectional study was conducted among 105 resident physicians working at 4 clinical departments of Peking Union Medical College Hospital in December 2014. The contents of the investigation included the cognition of ultrasound examination ，the communication with ultrasound physicians and the role of ultrasound in clinical practice. Results A total of 100 valid questionnaires was gained. 81% of the resident physicians understood the ultrasound indications slightly or moderately, and the results were moderately or quite approved by 91% of residents. 70% of resident physicians had a slight or moderately understanding for bedside ultrasound indications.90% of the physicians thought that ultrasound is quite and extremely important in clinical work,while 59% have an overall medium satisfaction rate for the interpretation of ultrasound report. Conclusion Explicit the needs of clinicians and training targeted will benefit the use of ultrasound in clinical work.

Development and implementation of an objective structured clinical examination (OSCE) in clinical medicine "7 to 8 transferring project" admission examination

2016, 36(1):  138-140. 

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Objective：To recruit top students from the medical institutions who completed six year education in a 7-year curriculum system at those schools in 8-year curriculum system at PUMC. Methods：Developing and implementing a 5-stations OSCE in clinical medicine "7 to 8 transferring project" admission examination. Results：Analyzing the test scores showed that, total score of OSCE/medical theory test/communication station shows a near-normal distribute. However history taking/physical examination/English test is not able to distinguish the difference. The correlation coefficient of t of each test station is between 0.048～0.334. Conclusion: It is necessary to develop and implement an OSCE for recruiting top medical students, and OSCE is proved a reliable tool for testing clinical ability.

The development and application of postgraduate degree administration system in medical college

2016, 36(1):  141-144. 

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As one of the sub-modules of the postgraduate administration system, development and application of the postgraduate’s degree administration system (PDAS) will optimize the working process, modernize administration and improve the efficiency. This article briefly introduced the design principles, classified authorization and main functional modules of such system, and summarized the features and benefits of its application.