Table of Content

05 March 2015, Volume 35 Issue 3

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RNAi induced integrin-linked kinase silencing in Tb human tongue cancer cells inhibits the growth of xenograft tumor

2015, 35(3):  289-294. 

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Objective: To investigate the effects of silencing integrin-linked kinase in Tb human tongue cancer cell on the expression and phosphorylation of its downstream signal molecular Akt and GSK3β and growth and spontaneous metastasis of tumor xenografts. Methods: Target cells were constructed by transfecting specific siILK plasmid and a non-homologous vector negative control into Tb cells. Tb, Tb vector and Tb siILK cells, respectively, were injected into nude mice subcutaneously．Weight and size of the xenograft tumor in nude mice were measured．Morphology of tumor tissue was observed by routine pathology methods．Changes in the expression and phosphorylation of Akt and GSK3β and micro-blood vessels in tumor tissue were detected by immunofluorescence and laser scanning confocal detection, respectively．Results: Phosphorylation of Akt and GSK3β in vivo and vitro was obviously inhibited in Tb siILK group compared with the other groups. Compared with Tb ［(3.58±1.04) g］and Tb vector ［(3.64±0.65) g］group, the mean tumor mass in Tb siILK group ［(1.68±1.35) g］decreased 53.0% and 53.8%，respectively (P<0.05)．Microvessel formation was also reduced in Tb siILK group compared to the other groups （P<0.05）. Conclusion: Silencing ILK inhibits proliferation and microvessel formation of Tb human tongue cancer tumor xenograft, which may be related with the inhibition of phosphorylation of Akt and GSK3β. The results suggest that ILK could be a therapeutic target protein for tongue cancer.

EPCs expressing hypoxia inducible factor 1αmu enhanced osteogenesis and angiogenesis of BMSCs transfected with bone morphogenetic protein 2

2015, 35(3):  295-302. 

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Objective To study the effect of endothelial progenitor cells (EPCs) expressing hypoxia inducible factor 1αmu on osteogenesis and angiogenesis of bone morphogenetic protein 2 transfected bone marrow stromal cells(BMSCs) .Methods BMSCs and EPCs were obtained and identified from rabbit bone marrow by density gradient centrifugation.To define the best ratio of EPCs and BMSCs,MTT assay was used to detect the proliferation of different proportion of BMSCs/EPCs.Experiment was divided into 6 groups according to different cells and genes transfected：group A：BMSCs ；group B：Adv-BMP2-BMSCs；group C：BMSCs plus EPCs；group D：Adv-BMP2-BMSCs plus EPCs；group E：BMSCs plus Adv-HIF1αmu-EPCs；group F：Adv-BMP2-BMSCs plus Adv-HIF1αmu-EPCs. Adv-HIF1αmu and Adv-BMP2 were respectively transfected into EPCs and BMSCs with the optimum multiplicity of infection (MOI).The expressions of BMP2 and HIF-1α proteins were detected by Western blot .The osteogenesis and angiogenesis potential of co-cultured cells were detected by alkaline phosphatase (ALP) activity and qRT-PCR.Result The co-culture proportion of BMSCs and EPCs was 1:1.Western blot showed that expression of BMP2 protein in groups of Adv-BMP2-BMSCs、BMSCs was higher than that in the groups of Adv-HIF1αmu EPCs and EPCs(P<0.05),expression of HIF-1α protein in group of Adv-HIF1αmuEPCs was higher than that in the groups of Adv-BMP2 -BMSCs、BMSCs and EPCs(P<0.05).Alkaline phosphatase activity and osteogenesis and angiogenesis related gene expression in group F were higher than that in other groups (P<0.05).Conclusion Adv-HIF1αmu EPCs co-cultured with Adv-BMP2 -BMSCs can promote osteogenesis and angiogenesis of Adv-BMP2-BMSCs.

Effect of Ginsenoside Rg3 on the proliferation, adhesion, migration and apotosis of human hepatoma cell lines and the mechanism

2015, 35(3):  303-307. 

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Object To investigate the effect of Ginsenoside Rg3 on proliferation, adhesion, migration and apoptosis of human hepatoma cell lines and reveal its mechanism. Methods The human hepatocellular carcinoma cell line HepG2, QGY and non tumor cell line HEL cells were used. By using the MTT test, cell adhesion assay, Transwell and flow cytometry, the effect of Rg3 on hepatocellular carcinoma cell proliferation, adhesion, migration and apoptosis were observed. The expression of CD44 and vascular endothelial growth factor(VEGF) protein were detected by Western blot after HepG2 and QGY cells exposed to Rg3. Results Rg3 could specifically inhibit the cell proliferation, adhesion capacity, migration of HepG2 and QGY cells and significantly induced the apoptosis (P<0.05). The expression of CD44 and VEGF protein were significantly down-regulated after Rg3 exposed to HepG2 and QGY hepatocarcinoma cells (P?0.05). Conclusion Rg3 inhibits cell proliferation, adhesion, migration of human hepatoma cell lines and significantly induce the cell apoptosis, and these effects are closely related with the expression inhibition of CD44 and VEGF protein.

Relationship between LCAT DNA methylation and serum lipips level in Hui patients with atherosclerosis in Ningxia

2015, 35(3):  308-312. 

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Objective To analysize the change of LCAT DNA methylation level and the relationship with blood lipids in Hui patients. Methods The 35 AS patients and 42 controls from Hui ethnic were confirmed by carotid artery Doppler ultrasound, LCAT mRNA expression were determined by real-time reverse transcription–polymerase chain reaction (qRT-PCR). The methylation status of LCAT gene promoter region was studied using the nest tounchdown methylation specific polymerase chain reaction (nt-MSP). The serum lipids were measured by ADVIA 2400 Automatic Biochemistry Analyzer. Results The TC,TG and LDL were higher than the normal group, HDL were lower than the normal group. Besides, compared with the normal group, the LCAT mRNA level of AS patients decreased by 61.2%(P＜0.01), and the LCAT DNA methylation level increased 1.3 times(P＜0.05). Conclusion The LCAT DNA hypermethylation is related to the occurrence and development of AS and the changes of serum lipids in Hui patients.

Ribonuclease inhibitor interacts with angiogenin and its mutant

2015, 35(3):  313-317. 

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Objective TO research the protein-protein interaction between ribonuclease inhibitor(RI) a?nd angiogenin(ANG) mutant. Methods Construct pCMV-3×flag-ANGH13R and pCMV-3×flag-ANGH114R pl?asmid.Immunofluorescence assay and laser scanning confocal microscopy analysis were used to observe the co-localization between RI and ANG mutant in BIU-87 cells.The interaction between RI and ANG mutant in BIU-87 cells was identified by glutathione-S-transferase (GST) pull-down assay and co-immunoprecipitation assay. Results The expressing plasmid pCMV-3×flag-ANGH13Rand pCMV-3×flag-ANGH114Rwas verified by restriction enzyme digestion and DNA sequencing.We found that RI was coloc-alized with ANG mutant in BIU-87 cell by laser scanning confocal microscopy analysis.Prokaryotically expressed products were purified and their sequences were confirmed.Glutathione-S-transferase(GST) pull do?w?n and co-immunoprecipitation results showed that the interaction between RI and ANG mutant occur?r?ed in BIU-87 cells. Conclusion The interaction between RI and ANG occurs in BIU-87 cells. The difference between RI and ANG mutant exists in BIU-87 cells.

Melatorin alleviates gastric mucosal injury induced by water-immersion and restraint stress in rats

2015, 35(3):  318-322. 

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Objective To investigative the protective effects of melatonin (MT) in gastric stress ulcer of rat and expression of MMP9, MMP2 and TIMP-2 in mucosa. Methods Stress ulcer model was established by water-immersion and restraint stress (WIRS). Melatonin at dose of 10、20、40、60mg/kg were administrated in different group of rat at 30 min before the onset of WIRS. The rat mucosa was examined and ulcer index were evaluated after 3 hours stress. Mucosa hemorrhage was determined at histology. In addition, the mRNA and protein level of MMP9、MMP2 and TIMP-2 was measured by PCR and western blot. Results The rat stress ulcer model can be established by WIRS at 3 hours. Pre-administration of melatonin can decrease the ulcer index (UI) and inflammatory cells in mucosa（P ＜ 0. 01). In addition, the mRNA and protein level of MMP9 and MMP2 was increased, TIMP-2 was decreased in WIRS induced rat ulcer model (P ＜ 0.01). Moreover, the mRNA and protein level of MMP2 and MMP9 was inhibited, while TIMP-2 was promoted by melatonin in stress induced ulcer（P ＜ 0. 05 or P ＜ 0. 01). Conclusions These results reveal the alleviation role of Melatonin on WIRS-induced gastric mucosal injury in rats, which may partly through regulating the expression of MMP2, MMP9 and TIMP2.

The effect of bFGF on the expression of NMDA receptor in spinal cord of rat with neuropathic pain

2015, 35(3):  323-328. 

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Objective To investigate the role of basic fibroblast growth factor (bFGF) and its effect on the expression of N-methyl-D- aspartate receptor in rat spinal cord with neuropathic pain. Methods Forty-eight male SD rats were randomly divided into four groups (n=12): sham group (group A), sham + bFGF antibody group (group B), neuropathic pain group (group C), neuropathic pain + bFGF antibody group (group D). Neuropathic pain model was produced by spared nerve injury (SNI). bFGF antibody 18 ug (40 ul) was injected intrathecally on 1, 6, 9, 13, 16 and 20 days after operation in group B and D. The equal volume of phosphate buffered saline was injected intrathecally in group A and C at the same time. The paw withdrawal mechanical threshold (PWMT) was measured on 1, 4, 7, 14 and 21 days after operation and 1 day before operation. Rats in each group were sacrificed on 21d after operation and the lumbar segments of the spinal cord (L4~6) were collected. The expression level of glial fibrillary acidic protein (GFAP) was detected by immunohistochemical analysis; The expression of N-methyl-D-aspartate receptor 1 subunit (NR1), N-methyl-D-aspartate receptor 2 subunit (NR2A or NR2B) were detected by RT-PCR and Western blot. Results Compared with group A and B, the level of PWMT was decreased and the expression of NR1 and NR2B in spinal cord was increased in group C and D (P<0.05). Compared with group C, the level of PWMT was increased and the expression of NR1 and NR2B in spinal cord was decreased in group D (P<0.05). Conclusion bFGF can reduce degree of neuropathic pain in rat with neuropathic pain induced by SNI, and the effect of bFGF on expression of NMDA receptor may associated with this phenomenon.

Prokaryotic expression，purification and functional identification of RGD-recombinant SAK- a1M fusion protein

2015, 35(3):  329-335. 

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Objective Our study aims to construct a recombinant prokaryotic expression plasmid of RGD-rSAK- a1M in E.col BL21. Furthermore, it aims to purify the fusion protein and determine its bioactivity. Methods The target fusion gene, RGD-rSAK-a1M，was amplified by overlapping PCR and inserted into a prokaryotic expression vector PEGX-6P-1 with a His-GST tag to construct a recombinant expression plasmid PEGX-6P-1- RGD -rSAK- a1M, which had a high efficiency of prokaryotic fusion-expression. E.coli BL21 was used to transform the recombinant plasmid and the expression of fusion protein was induced by IPTG. The tag of His-GST was excised by a 3C protein after the purification through the Nickel ion affinity chromatography column, and the fusion protein was purified by a DEAE ion exchange column and molecular sieve. Soluble fibrin plate methods and inhibition test of platelet aggregation were applied to determine and evaluate its fibrinolytic activity and anti-platelet aggregation, respectively. Results 1) The fusion protein of RGD-rSAK- a1M was construted successfully. 2) The high express of target fusion protein in the supernatant of E.coli lysate was achieved, and purify the fusion protein. 3) Vitro experiments indicated that the fibrinolytic activity of purified fusion protein was basically the same as that of urokinase standards. 4) The effect on anti-platelet aggregation was significantly enhanced by fusion protein rather than simple recombinant staphylokinase. Conclusions Literature retrieval confirm that the soluble fusion protein of RGD -rSAK- a1M is firstly acquired, which has an equal fibrinolysis activity and a higher ability of anti-platelet aggregation compared to urokinase standards, and the ?effect on anti-platelet aggregation was significantly enhanced rather than simple recombinant staphylokinase，laying the foundation for following identification of fusion protein immunogenicity.

Calpeptin inhibits the proliferation of ER negative MDA-MB-231 breast cancer cells induced by EGF

2015, 35(3):  336-340. 

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Objective To investigate whether calpain is involved in epidermal growth factor (EGF)-enhanced colony formation in estrogen receptor (ER) negative breast cancer cells. Methods ER-negative MDA-MB-231 breast cancer cells were employed as a model system, Western lot analysis was used to observe protein alterations. Colony formation assay applied to evaluate the role of calpain in EGF-stimulated cell growth. Calpain specific inhibitor, calpeptin (Calp), was employed for pre-treatment of cells to observe its effect on the EGF actions. Results (1) EGF induced over-expression and truncation of FAK (also of cyclin E) protein in MDA-MB-231 breast cancer cells（p<0.01）; (2) EGF-enhanced processing of FAK could be blocked by calpeptin（p<0.05）; (3) EGF induced calpain 1 autolysis and this effect could be tremendously suppressed by pre-treatment with calpeptin（p<0.05）, and (4) Calpeptin greatly inhibited EGF-enhanced cell growth（p<0.05）. Conclusion Calpain is possibly a potential target for the treatment of ER-negative breast cancer.

Molecular expression of RAS components in heart and kidney of db/db mice

2015, 35(3):  341-344. 

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Objective To investigate the type 2 diabetes-induced expression changes of renin-angiotensin system (RAS) components in heart and kidney. Methods The db/db mice were applied to be the type 2 diabetes animal model. Gene and protein expression of RAS components in heart and kidney were measured by RT-PCR and immunoblotting, respectively. Results The mRNA expression of AGT (P < 0.01) and the protein expression of Renin (P < 0.01) and Ang II (P < 0.01) were significantly increased in heart of db/db mice, moreover, the mRNA (P < 0.001) and protein (P < 0.01) expression of Renin and the protein expression of Ang II (P < 0.05) were significantly up-regulated in kidney of db/db mice. Conclusion Type 2 diabetes induced the abnormal expression of RAS components in heart and kidney, consequently, leading to the increase of production of RAS active peptide Ang II.

XAV939 inhibits vasculogenic mimicry formation in human hepatoma hepG2 cells

2015, 35(3):  345-349. 

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Objective To investigate the impact of XAV939 to human hepatoma HepG2 cells vascular mimicry and its possible mechanism. Methods HepG2 cells were divided into control group and XAV939 treatment groups with dose of 0.5 and 1μmol/L; Tube formation of tumor cells experiment to detect the forming tubes ability of each group , RT-PCR were used to detect expressed ZEB1 and MMP-7 mRNA levels , Western blot were used to detect expression of β-catenin, ZEB1 and MMP-7 on protein levels. Results The number of vascular mimicry in the control group and the experimental group. In the control group : 9.67 ± 0.70 , 0.5μmol/L group : 5.67 ± 0.64 , 1μmol/L group : 2.27 ± 0.81,which means the number of pipeline structure formation reducing in vitro (P<0.01). Experimental group compared with the control group, ZEB1 and MMP-7 mRNA expression reduce; ZEB1, MMP-7 and β-catenin protein expression decreased(P<0.05). Conclusion XAV939 can effectively inhibition of vasculogenic mimicry formation in human hepatoma HepG2 cells.

Relationship of the markers and proliferation in vitro of circulating endothelial cells from chronic myelogenous leukemia patients

2015, 35(3):  350-354. 

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Objective To investigates the relationship of markers and proliferation in vitro of CECs from CML patients. Methods CECs were sorted by MACS and cultured to be compared with HUVECs in biology feature. We also examined expression of bcr/abl fusion gene by fish in 12 CML patients. Results The cultured CECs and HUVECs had typical cobble-stone morphology; they could be identified by high level of CD146 and VWF. CECs derived from CML patients have higher level of CD34、KDR and CD133, and showed more increased proliferative potential as compared with HUVECs(P <0.05). In 12 CML patients, the positive rate of bcr/abl fusion gene in CECs was 10.77%. Higher expressions of bcr/abl fusion gene have positive correlation trend with progression of disease. Conclusion CECs in patients with CML were different from mature endothelial cells, they gained more increased proliferation by carrying bcr/abl fusion gene.

The effect of trimetazidine and/or rosuvastatin on improving the cardiac function damage of rats with myocardial infarction

2015, 35(3):  355-359. 

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Objective This study aimed to investigate the cardioprotection of trimetazidine and/or rosuvastatin in rats with myocardial infarction (MI). Method MI was introduced to SD rats by ligating the left anterior descending coronary. Rats were randomly assigned into following groups: sham group, Mi group, trimetazidine (T) + MI group, rosuvastatin (R) + MI group and T + R + MI group. After treatment for 14 days, all groups' hemodynamic parameters were measured. Apoptosis index (AI) were detected by TUNEL staining. The myocardial infarction area was measured by Evans/TTC staining. The expression of bcl-2 and bax was determined by western blot assay. Results When compared with sham group, the LVSP, dP/dTmax and dP/dTmin of MI rats significantly reduced (P<0.01), but LVEDP markedly increased (P<0.01). When compared with the MI group, the LVSP, dP/dTmax and dP/dTmin in the T + MI group, R + MI group and T + R + MI group significantly increased, and the LVEDP dramatically reduced (P<0.05). The apoptosis index in the MI group significantly increased accompanied by marked increase in the expression of Bcl-2 and Bax, especially the Bax when compared with sham group. After treatment with T and R, the apoptosis index reduced and the expression of Bcl-2 and Bax decreased significantly, mainly the Bax. The above changes were the most evident in the T+R+MI group. Conclusion Treatment with T and/or R can improve the heart function of MI rats, which may be attributed to the reduction in the apoptosis index and decrease in the expression of Bax and Bcl-2 in the myocardium. The cardioprotection is the most obvious after treatment with T and R suggesting the synergistic effect.

The quantity and phenotype change of regulatory T cell in the model of mice liver transplantation

2015, 35(3):  360-365. 

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Objective To research the degree of immune graft rejection, the quantity and function of regulatory T cells in the model of liver transplantation after mice liver transplantation to provide theory evidence in revealing mechanisms of immune tolerance. Method Transplantation liver samples were collected at 3d, 7d, 14d, 28d after allogeneic or syngeneic mice liver transplantation (N=4 for each group at one time point). Graft rejection score was given with HE staining. T and B lymphocytes and change of regulatory T cells were identified with IHC staining.The change of Foxp3 mRNA and CLAT-4 on surface of regulatory T cells were detected by real-time PCR and IFC respectively. Results Allogeneic graft was rejected during 2w after transplantation, and the rejection receded generally after 2w. Inflammatory cells intra-graft was mainly T lymphocytes. Foxp3 positive cells and mRNA of Foxp3 in allogeneic graft group was significantly higher than these in isogeneic graft group (15.0+1.3 vs. 2.7+1.0，P<0.05）. Meanwhile, expression of CLAT-4 on regulatory T cells also was higher in allogeneic graft group compared with isogeneic graft group(53%+3% vs. 13%+2%,P<0.05). Conclusion The immunity rejection after allogeneic liver transplantation achieved by regulatory T cells negative regulation and then achieved immune tolerance.

Protein expression pattern and correlation with clinicopathological features in clear cell renal cell carcinoma neural Wiskott–Aldrich syndrome

2015, 35(3):  366-370. 

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Objectives To examine the level of N-WASP expression in clear cell renal cell carcinoma (CCRCC) and its association with clinicopathological features. Methods Immunohistochemical staining for N-WASP was performed on tissue microarrays constructed from tumor and para-tumor tissue obtained from 73 CCRCC patients. The difference in N-WASP expression between tumor tissue and adjacent normal renal tissue was examined. Correlations between N-WASP expression in the tumor and clinicopathological parameters were analyzed and the relationship between N-WASP expression and overall survival also assessed. Univariate and multivariate survival analyses were performed using the Cox proportional hazards regression model. Results N-WASP expression was reduced in tumor tissues (P<0.001). Patients with weak-to-moderate N-WASP expression tended to have a longer over-all survival than those with a high tumor N-WASP level (P<0.01). A higher level of N-WASP expression in the tumor was an indicator of poor survival in CCRCC patients (P<0.01). N-WASP was an independent predictor for overall survival (P<0.05). Conclusions N-WASP was significantly downregulated in CCRCC and could serve as a promising prognostic biomarker for predicting clinical outcome.

Overexpression of GSK-3β promotes the proliferation and invasion of human glioma cell

2015, 35(3):  371-376. 

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Objective To explore the expression levels of Glycogen Synthase Kinase-3β (GSK-3β) in glioma cells and its function on cell proliferation and invasion. The underlying mechanism was also investigated. Methods The expression levels of GSK-3β in four glioma cell lines (A172, T98, U87, U251) and normal human astrocytes 1800 were analyzed by RT-PCR and Western blotting. The constructed pcDNA3.1(+)-GSK-3β（rGSK-3β）recombinant vector and PAX3 siRNA were transfected into U87 cells. Cell viability and invasion ability were detected by MTT and Transwell invasion assay, respectively. Results The GSK-3β expression levels were detectable in four glioma cell lines （P<0.05）, but low in the control 1800 cells. Overexpression of GSK-3β in U87 cells increased rates of cell proliferation was assessed by MTT assay and Ki67 expression. The GSK-3β expression promoted cell invasion（P<0.05）. Further analysis suggested that the enhanced PAX3 was responsible for GSK-3β-induced cell proliferation and invasion. These advantages were abolished by inactivation of PAX3 using specific PAX3 siRNA (P<0.05). Conclusion This research demonstrated that GSK-3β, via PAX3, promotes the rates of cell proliferation and invasion in U87 glioma cells.

Lidamycin induces cell apoptosis and cell cycle arrest of human multiple myeloma RPMI 8226 cells

2015, 35(3):  377-382. 

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Objective To investigate the effect of lidamycin (LDM) against multiple myeloma and its mechanism. METHODS The human multiple myeloma RPMI 8226 cells in logarithm growth phase were selected and were randomly divided into control group, 0.1, 0.5 and 1 nmol/L LDM groups. MTS assay was used to detect the proliferation rate of RPMI 8226 cells and the flow cytometry method was used to analyze the distribution of cell cycle and cell apoptosis in various groups. The expression levels of protein associated with apoptosis and cell cycle were detected by Western blotting method. Results After 48 h cell culture, the cell proliferation rate in LDM groups was lower than those in control group (P < 0.05). The number of S phase and G2/M phase cells in LDM groups was higher than those in control group (P < 0.05). The cell apoptosis rate in LDM groups was higher than those in control group (P < 0.05). The expressions of cleaved caspase-3, caspase-7, caspase-9 and poly ADP-ribose polymerase (PARP) in LDM group were higher than those in control group (P < 0.05). The expressions of p21 and p27 in LDM group were higher than those in control group (P < 0.05). Conclusion LDM can induce apoptosis by increasing the levels of cleaved caspase-3, caspase-7, caspase-9, and PARP in cells and LDM can induce the S phase and G2/M phase arrest by increasing the levels of p21 and p27 in human multiple myeloma RPMI 8226 cells.

Effect of Runx3 on the proliferation, apoptosis and invasion of HepG2 cells

2015, 35(3):  383-386. 

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Objective To explore the effect of siRNA targeting Runx3 on proliferation, apoptosis, and invasive ability of HepG2 cells. Methods The Runx3-siRNA was transfected into HepG2 cells. HepG2 cells were divided into four groups: control group, liposomes group, NC-siRNA group and Runx3-siRNA group. The expression levels of Runx3 mRNA and Runx3 protein were examined by RT-PCR and Western blotting. MTT assay, flow cytometry and Transwell chamber were used to determine the effect of Runx3 on HepG2 cells proliferation, apoptosis and migration. Results MTT assay showed that the proliferation of HepG2 cells was obviously inhibited by Runx3-siRNA. The result of flow cytometry demonstrated that the apoptosis of HepG2 cells was promoted by Runx3-siRNA. The invasion of HepG2 cells was also significantly inhibited by Runx3-siRNA according to Transwell chamber assay. Conclusions Silence Runx3 could effectively inhibit cell proliferation and migration, and promote apoptosis in HepG2 cells. Our study shows that Runx3 gene plays an important role on the occurrence and development in hepatocellular carcinoma. Runx3 could be a new target for the treatment of hepatocellular carcinoma.

The role of histone deacetylase inhibitor largazole prevent aging of hUCMSCs

2015, 35(3):  387-392. 

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Objective This research adopt the histone acetylation inhibitor Largazole to treat on human umbilical cord mesenchymal stem cells (hUCMSCs), and discuss the role and molecular mechanism of anti-aging. Methods hUCMSCs were isolated from human Umbilical cords after trypsin digestion, and determined by Flow cytometry analysis. they were cultured with different concentrations of largazole to find the fine cell proliferative concentrations. Passage 4, 8, 12, and 16 cells were subjected to real-time PCR for the change of stem cell gene expression on largazole group. We investigated the effect of largazole, against the histone H3 lysine 9 or 14 (H3K9/K14) acetylation of the gene (TERT, OCT4 and OPN) by CHIP assays. Results hUCMSCs can undergo spontaneous differentiation and aging during expansion. Low concentrate largazole can promote hUCMSCs proliferation and suppresse its spontaneous osteogenic differentiation. Largazole modulates histone H3 acetylation in the TERT, OCT4 and OPN genes; Conclusion our results indicate that histone H3 acetylation, which can be modulated by Largazole, plays a key role in regulating hUCMSCs biological characteristics.

The expression and significance of HIF-1α and HIF-2α in different tissues for plateau microtus oeconomus

2015, 35(3):  393-394. 

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Testosterone inhibits macrophage-derived foam cells formation in mice

2015, 35(3):  395-396. 

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Regulation of lovastatin on NO in neonatal rat cardiac fibroblasts in vitro induced by aldosterne

2015, 35(3):  397-398. 

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Sinomenine down-regulates adiponectin and expression of it’s receptors in rabbit knee osteoarthritis model

2015, 35(3):  399-400. 

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Research progress of autophagy and tumor

2015, 35(3):  401-404. 

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Autophagy has a dual role of suppression and promotion in tumor genesis and progression, and autophagy can dictate the development direction of some tumor. It have achieved tumor treatment effect by activating or inhibiting autophagy. So it may become a new target for tumor therapy by regulating autophagy.

Advance of mitochondria SIRT5-regulated metabolic protein post-translation modification

2015, 35(3):  405-408. 

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SIRT5, a member of Sirtuin protein family, is mainly located in the mitochondria and involved in multiple forms of protein post-translation modification, such as deacetylation, desuccinylation and demalonylation, regulating energy metabolic process, cellular redox reaction and apoptosis and thus shed light upon the basic function of SIRT5.

The central role of GDNF-regulatory network in the self-renewal of mammalian spermatogonial stem cells

2015, 35(3):  409-412. 

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Glial cell line-derived neurophic factor(GDNF) is secreted by sertoli cells, peritubular myoid cells, spermatocytes and roud spermatids in testes, is a critical component during self-renwal of spermatogonial stem cells(SSCs) self-renewal. GDNF binds to GFRα1, expressed on the surface of SSCs, to form a heterodimer complex. As a consequence, this complex via activation of RET, triggers MAPK, SFK and PI3K-ATK signaling pathway to regulate SSCs self-renewal and differentiation.

The effect of nitric oxide on neuronal apoptosis during cerebral ischemia-reperfusion

2015, 35(3):  413-416. 

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Nitric oxide level of cerebral tissue was "double peaks-like" during cerebral ischemia-reperfusion, and the role of nitric oxide will change depending on its level in vivo. Nitric oxide mediated apoptosis mainly through four signal pathways. Nitric oxide can make related protein s-nitrosylation via molecular modification, further regulate signal pathways and affect apoptosis.

Advances in the research of targeting the signaling pathways of leukaemia stem cells in chronic myeloid leukaemia

2015, 35(3):  417-420. 

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Leukaemia stem cells (LSCs) is the main source of drug resistance, recur- rence and poor prognosis in treatment of chronic myeloid leukemia (CML). Specific elimination of LSCs is expected to become a way to cure CML. The signaling path-ways in LSCs are abnormal, such as WNT/β –catenin,JAK/STAT,PI3K/AKT, and so on.

Advances in the researches of the therapeutic HPV DNA vaccines

2015, 35(3):  421-424. 

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HPV infection is closely related to cervical cancer.Therapeutic HPV DNA vaccines can treat HPV infection so that they can prevent cervical intraepithelial neoplasia which is caused by HPV.So we mainly discuss about HPV,therapeutic HPV DNA vaccines,improvements and so on.In order to be able to be applied in clinical in the future,researchers use different methods to improve the therapeutic HPV DNA vaccines which is based on intensive study of HPV and DNA vaccines.

Research progress of molecular markers in bladder urothelial carcinoma

2015, 35(3):  425-428. 

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Tumor markers as a non-invasive examination method not only play very important role in the diagnosis and classification of cancer, also has an important role to monitor disease recurrence and determine the efficacy of treatment.At present, the existing methods do not achieve the desired detection sensitivity and specificity. With the development of high-throughput technology, many molecular markers for bladder cancer have been discovered , such as DNA / RNA, proteins, and metabolites or the combination of them provide a more accurate methods for bladder cancer detection.

Exploration and practice in the reform of the grant system of medical postgraduate

2015, 35(3):  429-431. 

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According to the related regulations, PUMCH has established the postgraduates grant system with special features of medical education. During the application and the implement of this system, some problems are revealed and some adjustments are made to improve it accordingly. On the basis of PUMCH’s management practices of grant system, the author analyzed and summarized the related features and difficulties，and made some recommendations for future improvement.

Establishing quality management system of innovation training project for college students

2015, 35(3):  432-434. 

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In order to improve the quality of innovation training project for college students. Pecking Union Medical College have investigated implementation effects of the project through questionnaires and made a thorough inquiry for project issues. And put forward improvements: revving up publicity and improving students participation, setting up information management model and enhancing awareness of process management, making dynamic management and building exchange platform, making the system of rewards and penalties better and enhancing students’ sense of achievement.