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Table of Content

    05 March 2014, Volume 34 Issue 3
    Designing of a mimic peptide based on Sinonatrix percarinata PLI γ sequence and determination of its inhibition activity
    2014, 34(3):  289-294. 
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    Objective To design a mimic peptide based on Sinonatrix percarinata PLIγ sequence and to explore its inhibitory activity on PLA2. Methods Sinonatrix percarinata PLIγ gene was cloned by RT-PCR and aligned with other published PLIγ sequences. Based on the results, a 19aa peptide was designed derived from Sinonatrix percarinata PLIγ. Molecular docking was then achieved to investigate the interaction between the 19aa peptide and sPLA2 by Auto-dock software. Experimental validation was conducted by agarose plate method, and anti-hemorrhagic activity against snake venom sPLA2 on mice skin was tested as well. Arachidonic acid content in LPS induced Raw264.7 cells was assayed to evaluate the 19aa peptide inhibitory effect on mammal sPLA2 enzyme. Results The mimic 19aa peptide with the sequence as PGLPLSYPNGGGGSVAFRS, behaved dominant pharmacological inhibitive properties to enzymatic activity and hemorrhage toxicity of sPLA2. It also significantly reduced arachidonic acid content in the LPS induced Raw264.7 inflammatory cells with an IC50 of 86.3μmol/L, probably by inhibiting the mammal sPLA2. Conclusion The designed 19aa mimic peptide showed similar pharmacological properties with natural Sinonatrix percarinata PLIγ, indicates its anti-inflammatory and anti-hemorrhage potential.
    Erythropoietin attenuates Cardiomyocytes hypertrophy induced by Ang Ⅱ and its possible signal pathway in rats
    2014, 34(3):  295-300. 
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    Objective To observe the effects of erythropoietin ( EPO) on angiotensinⅡ ( AngⅡ ) induced neonatal rat cardiomyocyte hypertrophy and its signal protein expression. Methods Cardiomyocytes were isolated from new-born Sprague-Dawley rats and were used to establish the model of hypertrophy by AngII in vitro.The cells was treated with EPO or/and P38MAPK inhibitor SB203580. The cell surface area, [3H]-leucine incorporation ,mRNA expression of atrial natriuretic factor(ANF) and β-myosin heavy chain(β-MHC) of cardiomyocytes were employed to detect cardiomyocyte hypertrophy. The mRNA levels of TGF-β1 and P38MAPK was analyzed by real-time quantitative PCR. The protein expression of TGF-β1、TAK1、P38MAPK and phosphorylation of TAK1 and P38MAPK were analyzed by Western blot. Results EPO attenuated hypertrophy of cardiomyocytes. EPO significantly suppressed TGF-β1、TAK1、P38MAPK、phosphrylation of TAK1 and P38MAPK expression. SB203580 represses cardiac hypertrophy. Conclusions EPO attenuates cardiomyocytes hypertrophy induced by Ang II,and it might be associated with TGF β1-TAK1-P38MAPK signaling pathway.
    Gene-gene interactions among VANGL1,FZD3 and FZD6 are associated with neural tube defects in Han population of Northern China
    2014, 34(3):  301-304. 
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    Objective To identify the impact of gene-gene interaction between VANGL1, FZD3 and FZD6 on the risk of neural tube defects in Han population of Northern China. Methods Two SNPs (rs4 839 469 and rs34 059 106) within VANGL1, two SNPs (rs2 241 802 and rs28 639 533) within FZD3 and three SNPs(rs827 528, rs3 808 553 and rs12 549 394) within FZD6 were genotyped in 135 NTD patients and 135 healthy controls.The gene-gene interaction between VANGL1, FZD3 and FZD6 was analyzed by Multifactor Dimensionality Reduction software. Results The frequency of genotypes of rs4 839 469 within VANGL1 and rs3 808 553 within FZD6 were significantly higher in patients than that in controls(P<0.05).MDR revealed that the significance were shown in patterns with 2 SNPs (rs4 839 469—rs3 808 553)(OR=3.18,95% CI:1.85~5.44;χ2=18.39,P<0.0001),3 SNPs( rs4 839 469—rs2 241 802—rs3 808 553) (OR=4.17,95% CI:2.43~7.14;χ2=28.5,P<0.0001)and 4 SNPs(rs4 839 469—rs2 241 802—rs827 528—rs3 808 553) (OR=7.34,95% CI:3.98~13.54;χ2=45.3,P<0.0001).Conclusion The gene-gene interaction between VANGL1, FZD3 and FZD6 may exist and increase the risk for NTDs.
    Proliferation And Androgen Receptor Phosphorylation Of Prostate Cancer Cells Induced By Epidermal Growth Factor At The Absence Of Androgen
    2014, 34(3):  305-309. 
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    Objective To investigate the influence of epidermal growth factor on the proliferation and androgen receptor phosphorylation of prostate cancer cells at the absence of androgen. Methods LNCaP and LAPC4 AR were used for the study, after EGF treatment,western-blot was used to detect AR phosphorylation in LNCaP and LAPC4 cells; Knockdown of Src and Ack1 genes with siRNA transfection to observe the effect on androgen receptor phosphorylation; Cell Counting Kit-8(CCK-8) was used to measure the cell proliferation; Quantitive RT-PCR was used to analyze the expression of prostate–specific antigen and human kallikrein-2 mRNA. Results EGF can induce AR specific site phosphorylation at Try-534 and Try-267 by Src kinase and another unknown kinase; Compared with untreated control, EGF improved the proliferation of prostate cancer cells(p<0.05) and increased the expression of prostate –specific antigen(p<0.05) and human kallikrein-2(p<0.05) Conclusions EGF can induce proliferation and AR phosphorylation of prostate cancer by intracellular non-receptor tyrosine kinases.
    The impact of miR-146a in breast cancer on the function of tumor-associated macrophage
    2014, 34(3):  310-313. 
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    Objective To investigate the influence of miR-146a on the function of tumor-associated macrophage (TAM), and the mechanism of its expression regulation. Methods We compared TAM in 4T1 xenografted tumor grown in BALB/c mouse to peritoneal macrophage (PEC), and TAM in human breast cancer to PBMC respectively to detect the expression level of miR-146a using q-PCR method. Mixed implantation subcutaneously of miR-146a antagomir-transfected macrophages with 4T1 cells was used to determine the function of miR-146a in TAM. Results miR-146a was significantly down-regulated in breast cancer-associated macrophage in both 4T1 tumor grown in mice (P<0.05) and breast cancer patients (P<0.01). It was observed that macrophage with decreased miR-146a expression inhibited the 4T1 tumor cell growth in vivo. Conclusion miR-146a can regulate tumor associated macrophage function, so as to promote tumor growth.
    Ubiquitin ligase MDM2 promotes the ubiquitination and degradation of exogenous NOLC1
    2014, 34(3):  314-317. 
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    Objective To study the role of MDM2 in the ubiquitination of NOLC1, a regulator of MDM2. Methods Full-length NOLC1 proteins and its nuclear localization signal region were cloned and expressed in E.coli. Recombinant human MDM2 with ubiquitin ligase activity was used to study the ubiquitination of NOLC1 by MDM2 in in vitro ubiquitination system. The degradation of exogenous NOLC1 by MDM2 was studied in mammalian cells. Results MDM2 promoted the ubiquitination of full-length NOLC1 proteins and its Nuclear localization signal region in in vitro ubiquitination system. MDM2 promoted the proteasome dependent degradation of exogenous NOLC1 by over 70% in mammalian cells. Conclusion Ubiquitin ligase MDM2 promotes the ubiquitination and degradation of exogenous regulator NOLC1, which provides important clues for study the regulation of MDM2-TP53-NOLC1.
    Emodin-Polylactic-Co-Glycolic Acid Nanoparticles delay chronic renal failure by 5/6 Nephrectomy
    2014, 34(3):  318-323. 
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    Objective To explore the kidney protecting effect and possible mechanism under which the renal tissues in 5/6 nephrectomy rats was intervened by emodin-polylactic-co-glycolic acid nanoparticles. Methods Rats were randomly divided into two groups, eight as Sham-operated group and twenty-four as model group. After rats model was successfully established, twenty-four rats in model group were randomly and equally assigned into Model group (Model group), emodin group eight(EMD)and nanoparticles group eihgt (NPs). Drugs were given to rats in different groups for a total 8 weeks, then 24h urine protein, blood urea nitrogen (BUN)and creatinine (SCr) were measured by their corresponding methods respectively; kidney pathological changes was observed under optical microscope; protein and mRNA expression of growth factor beta-1(TGF-β1) as well as connective tissue growth factor (CTGF) in renal tissues were detected respectively by RT-PCR methods and immunohistochemistry. Results After treated with drugs,compared with sham group,24h urine protein,blood urea nitrogen, serum creatinine, mRNA and protein expression of TGF-β1 and CTGF of rats in model group were both increased(P<0.01);Compared with model group,24h urine protein,blood urea nitrogen,serum creatinine, mRNA and protein expression of TGF-β1 and CTGF of rats in EMD group and NPs group were both decreased(P<0.01);Compared with EMD group,24h urine protein,blood urea nitrogen,serum creatinine, mRNA and protein expression of TGF-β1 and CTGF of rats in NPs group were both decreased(P<0.01).Conclusions EMD-PLGA NPs could thus play an effect of inhibiting renal fibrosis and slowing the progress of chronic kidney disease more obviously than ordinary emodin.
    Analgesic mechanism of lappaconitine in rat midbrain periaqueductal gray
    2014, 34(3):  324-327. 
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    Objective To investigate the analgesic effect of lappaconitine (LA) on burned rats and its supraspinal analgesic mechanism. Methods Male Sprague-Dawley rats were divided into 6 groups: normal, burn, burn LA,burn LA plus antisense oligodeoxynucleotide (A-ODN),burn LA plus A317491 and burn LA plus ATP. The later 4 groups had LA (6mg/kg) administered intraperitoneally. The pain threshold was characterized by the mechanical withdrawal threshold (MWT) testing; The expression of P2X3 receptor protein in the midbrain periaqueductal gray (PAG) was assessed by Western blot analysis. Observation the intra-PAG injection of P2X3 A-ODN, selective antagonist and agonist of P2X3 receptor on the analgesic effect of LA. Results In burn group, the MWT was decreased while increased in burn LA group. LA induced a significantly up-regulated P2X3 receptor protein expression in PAG. The antinociceptive effect of LA was attenuated significantly by P2X3 A-ODN. The α,β-meATP or A317491 significantly enhanced or attenuated analgesic effect of LA, respectively. Conclusions LA produces an antinociceptive effect on burn pain via increases expression and function of P2X3 receptors in the PAG.
    Effets of VPA combining with arsenic trioxide on the expressions of VEGF,NF-κB and MMP-9 in NB4 cell line
    2014, 34(3):  328-331. 
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    Objective To investigate the expressions of vascular endothelial growth factor、NF-κB and MMP-9 in NB4 cell lines affected by VPA in combination with arsenic trioxide. Methods The experimental included control group、the group of arsenic trioxide、the group of VPA and the group of arsenic trioxide combined with VPA, MTT assay method was used to detect the cell survival rate. RT-PCR was used to detect the mRNA level of VEGF、NF-κB and MMP-9.Western Blot was used to detect the protein level of VEGF、NF-κB and MMP-9. Results VPA can increase the decreased survival rate induced by arsenic trioxide in NB4 cell lines, the expression of VEGF、NF-κB and MMP-9 in NB4 cell lines were increased under the effect of arsenic trioxide (P<0.05). The expression of VEGF、NF-κB and MMP-9 in the group of VPA combined with arsenic trioxide were all decreased than in the group of arsenic trioxide alone(P<0.05).Conclusion VPA can down-regulate the increased expression of VEGF、NF-κB and MMP-9 caused by arsenic trioxide.
    The roles of RhoA/Rho kinase in hepatic fibrosis in rats with type 2 diabetes
    2014, 34(3):  332-338. 
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    Objective To determine whether RhoA/Rho-kinase is involved in the pathogenesis of hepatic fibrosis in rats with type 2 diabetes. Methods An animal model of type 2 diabetes was developed by high fat diet combined with intraperitoneal injection of low-dose streptozotocin (STZ, 30mg/kg, i.p.). Fasting blood glucose, triglycerides, cholesterol, aspartate aminotransferase and alanine aminotransferase was measured. The deposition of collagen in liver was evaluated by Masson staining and the hydroxyproline determination. The mRNA expressions of transforming growth factor (TGF-β1) and connective tissue growth factor (CTGF) in liver tissue were assessed by RT-PCR. Immunohistochemistry staining for TGF-β1 was performed. The phosphorylation of myosin phosphatase target subunit 1(p-MYPT1) was measured by Western blot analysis. Results Compared with control rats, Blood glucose, blood fat, aminotransferase and the deposition of collagen in the liver in the untreated diabetic rats was significantly increased (P<0.01); p-MYPT1 and TGF-β1 protein, the mRNA expression of TGF-β1 and CTGF was significantly enhanced (P<0.01). Compared with untreated diabetic rats, treatment with fasudil significantly decreased aminotransferase, deposition of collagen, p-MYPT1, mRNA expression of TGF-β1 and CTGF (P<0.01). Conclusions Hyperglycemia can activate the RhoA/Rho-kinase which maybe regulates TGF-β1/CTGF expression in liver tissues. RhoA/Rho-kinase plays an importent role in the development of diabetic liver fibrosis.
    The Runx3 gene methylation and protein expression in serrated lesion tissues
    2014, 34(3):  339-344. 
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    Objective To investigate the role and significance of methylation status and protein expression of the Runx3 gene in serrated lesions and carcinogenesis pathway. Methods The Runx3 gene promoters methylation were detected with the Taqman probe based real-time PCR(Methylight) technology in 77cases serrated lesions(29 cases of hyperplastic polyps, 29 cases of sessile serrated adenoma/polyp and 19 traditional serrated adenoma) , 16 cases of normal mucosa tissues and 14 cases of colorectal cancer. At the same time, immunohistochemical staining was utilized to detect the expression of Runx3, and to analyze the relationship of them. Results The rates of methylation of the Runx3 gene in normal mucosa tissues , HP, SSA/P, TSA and CRC were 12.5%(2/16)、17.2%(5/29)、51.7%(15/29)、63.2%(12/19) and 78.6%(11/14).The results of Immunohistochemistry showed that the positive expression of Runx3 in normal mucosa tissues , HP, SSA/P, TSA and CRC were 81.3%(13/16), 72.2%(21/29), 48.3%(14/29), 31.6% (6/19)and 21.4%(3/14). The Runx3 methylayion was correlated negatively with the expression of Runx3 protein in SSA/P, TSA and CRC (P<0.05). Conclusion Runx3 gene promoter region methylation induced Runx3 protein expression is cut or missing, the main reasons for the serrated lesions especially the occurrence of serrated adenoma and cancer pathyway plays an important role.
    The role and mechanism of ERK1/2 MPAK pathway in proliferation and migration of Eca109 cell line in vitro
    2014, 34(3):  345-349. 
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    Objective To investigate the role and mechanism of ERK1/2 MPAK pathway in regulating proliferation and migration of Eca109 cell line in vitro. Methods We treated Eca109 cells with MAPK(ERK1/2)inhibitor(U0126); cell counting method detected the cell proliferation; Cell morphology was observed under inverted microscope;qRT-PCR detected ERK1/2 mRNA level; Western blot determined the expression of ERK1/2 protein level; MTT and Wound scratch assay detected cell proliferation and migration; qRT-PCR examined miR-21 expression. Results Cell growth was inhibited by U0126 at 20μmol/L doses(p<0.05), destroyed the normal cell morphology; inactivited ERK1/2 MPAK pathway at protein level(p<0.05); inhibition of ERK1/2 MPAK pathway repressed cell proliferation and migration of Eca109 cells and downregulated the expression of miR-21(p<0.05). Conclusion Inhibition of ERK1/2 MPAK pathway weakened cell proliferation and migration of Eca109 cells, one of the most possible mechanism is associated it’s inhibition role on miR-21 expression.
    Construction of osteopontin siRNA expression lentivirus vector and itsinhibitory effect on the proliferation of SW480 cells
    2014, 34(3):  350-354. 
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    Objective To construct the recombinant lentivirus vector and explore its effects on colon cancer cell line SW480. Methods The siRNA interfering sequence targeting to osteopontin gene was designed and synthesized,then DNA segment was gained through annealing after chemosynthesis,and then was cloned to pSicoR vector, the recombinant lentivirus vector was constructed and used to challenge SW480 cells.The expression level of osteopontin mRNA and protein was detected by Real Time PCR and Western blotting. The effects on SW480 cells proliferation were analyzed by MTT assay. Results The restriction enzyme digestion,DNA sequencing and detection of GFP expression demonstrated that recombinant lentivirus vector was constructed successfully. Real Time PCR and Western blot analysis confirmed that the expression of OPN mRNA and protein were inhibited.The MTT assay showed that the anti-proliferation effect of lentivirus vector of osteopontin-siRNA on SW480 cells were significant(P<0.05). Conclusions The recombinant lentivirus vector could effectively inhibit the expression of osteopontin in SW480 cells and depress the proliferation of SW480 cells.
    Left ventricular myocardial strain in hypertensive patient with different geometric patterns using speckle tracking imaging
    2014, 34(3):  355-359. 
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    Objective To assess left ventricular systolic function with the peak systolic strain of longitudinal、radial and circumferential in patients with essential hypertension by using speckle tracking imaging . Methods 63 cases of patients with essential hypertension(EH) were enrolled, who were classified into the left ventricular hypertrophy group (n=33) and the non left ventricular hypertrophy group (n=30), according to the left ventricle mass index (LVMI). The peak systolic longitudinal strain (LS)、the peak systolic radial strain(RS) and circumferential strain (CS) were measurened in left ventricular long-axis and short-axis views by STI software. Results the peak systolis longitudinal strain of segments and global of left ventricular of group B decreased compared with the control group and group A(P<0.01); the peak systolic radial strain of three segments of hytension group A increased significantly composed with control group(P<0.01), but RS of group B slightly decreased compared with the control group and group A (P<0.01); the peak systolic circumferential strain of three segments of group B were slight lower than the control group and group A( P<0.05); Conclusion Speckle tracking imaging can assess left ventricular systolic function in gloal or local regiong of essential hypertention with different model, which were regard as a new method to evaluate early the left ventricular systolic function with essential hypertention.
    NF-E2-Related Factor2 Alleviated Non-Alcoholic Steatohepatitis in Mice
    2014, 34(3):  360-364. 
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    Objective To investigate the clinical significance of NF-E2-related factor2 in preventing and treating non-alcoholic steatohepatitis(NASH). Methods C57BL/6 mice were randomly divided into control group (control), model group (MCD) and Curcumin induced Nrf2 group (MCD+Curcumin). Mice in control group were fed with a standard diet while those in other two groups were fed with methionine choline-deficient (MCD) diet. At the same time, mice in Curcumin induced Nrf2 group were gavaged with 0.9 ml of curcumin daily. Control group and MCD group were gavaged with equal water daily. 4 weeks later, Nrf2 expression level, liver index, serum ALT and AST levels , serum triglyceride (TG), cholesterol (Chol) and GLU contents were measured, the HE and Oil Red O staining of liver cross section were observed, hepatic TG, Chol levels, MDA contents, GSH levels and SOD activity were quantified. Cytochrome c and ATP are also measured to investigate the mitochondrial function. Result After gavaged with Curcumin, compare to MCD group, the mice’s liver showed decreased lipid accumulation, serum ALT , AST, MDA, hepatic TG, Chol, and elevated serum TG and Chol levels.MDA level, Cytochrome c level also decrease, while GSH contents ,SOD activity and the content of ATP increase (P<0.05). Conclusion Nrf2 has the effect of protection of NASH from MCD diet and primarily through the way of reducing oxidative stress and increasing mitochondrial function.
    Method optimization of HIF-1αtransfection and its effects on survival rate and differentiation of mesenchymal stem cells
    2014, 34(3):  365-371. 
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    Object To investigate a method of MSCs transfection with HIF-1αgene-recombined liposome. Method HIF-1αwas transferred into the third generation MSCs by liposome. The expression of green flouresence protein (GFP) was observed and the efficiency of transfection was assessed. The three groups of cells, which are MSCs transfected with HIF-1αgene-recombined liposome, with control vector or with no transfection, were cultured in the hypoxia environment simulated through CoCl2. The expressions of HIF-1αmRNA, HIF-1αprotein of the different groups of MSCs were compared. Result pcDNA3.0-HIF-1α-eGFP could be transfected into MSCs by the liposome method successfully and the efficiency of the transfection is 21%. MSCs transfected with HIF-1α can be induced to differentiate to osteoblast and lipoblast successfully and HIF-1αtransfection may increase MSCs survival rate under hypoxia condition(P <0.05).The group of HIF-1α-transfected cells express much more HIF-1αmRNA and HIF-1α protein than the other two(P <0.05). Conclusion The study shows that the liposome can be used to transfect HIF-1α into MSCs successfully and MSCs transfected by HIF-1α gene-recombined liposome can strongly express HIF-1α mRNA and HIF-1α protein.
    Effect of N-cadherin on survival,proliferation and migration of neural stem cells transplanted into injured spinal cord of rats
    2014, 34(3):  372-375. 
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    Objective To investigate the effect on survival,proliferation and migration of NSCs transplanted into SCI of rats. Methods 60 SD rats were divided into control group, SCI group, transplanting group 1 and transplanting group 2. Using lentivirus virus as a vector, N-cadherin was transfected into primary cultured NSCs to implant in transplanting group 2. Neurological function were evaluated by BBB score. Immumofluorescence was used to detected the survival,proliferation and migration of NSCs at 5, 10 and 15d. Results BBB score of SCI group was lower than control group. BBB score in two transplanted groups increased significantly(P<0.05), with most evident in transplanting group 2(P<0.05). The number of Nestin-positive cells in transplanting group 2 was more than that in transplanting group 1 at the corresponding time point(P<0.05). The number of Nestin-positive cells at 15d was the most among three time points(P<0.05). The immigrating distance in transplanting group 2 was longer(P<0.05).Conclusion N-cadherin can promote the survival, proliferation and migration of NSCs in SCI rats to repair neurological function.
    Effect of CCN5 on proliferation, migration and invasion of hepatoma carcinoma cells and the mechanism
    2014, 34(3):  376-380. 
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    Objective To study the effect of CCN5 on proliferation, migration and invasion of hepatoma carcinoma cells and the mechanism. Methods Hepatocyte High Expression System of CCN5 was constructed and transfected HepG2 cells to make CCN5 overexpression, and then cell proliferation detection reagent box and Transwell chamber were used to determine the effect of CCN5 overexpression on proliferation, migration and invasion of HepG2. To further study the mechanism, phosphorylation levels of Smad2 and p44/42MAPK were determined. Results CCN5 was overexpressed in HepG2 cells. After that, the proliferation, migration and invasion of the CCN5 overexpression group were significantly inhibited (P <0.05) compared with the normal control group. Phosphorylation levels of Smad2 and p44/42MAPK were also significantly inhibited. Conclusion Overexpression of CCN5 can inhibit proliferation, migration and invasion of HepG2, which plays an important role in the formation of human hepatoma and proposes a new idea for the prevention and treatment of hepatic carcinoma.
    P38 MAPK inhibitor SB203580 decreases the AQP4 expression and cerebral edema of rats after cerebral ischemia and reperfusion
    2014, 34(3):  381-385. 
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    Objective To investigate the role of P38 inhibitor SB203580 on cerebral edema and Aquaporin 4(AQP4) expression after cerebral ischemia reperfusion in rats. Methods Fifty-four SPF male Sprague-Dawley rats weighing 240-260 g were randomly divided into 3 groups: sham operation group(sham grope); ischemia reperfusion group(I/R group); P38 inhibitor group(SB203580 group).The focal ischemia reperfusion rat model was established by the modified suture method. The rats of I/R group and SB203580 group were exposed to DMSO(10%,400μg/kg) or P38 inhibitor SB203580(400μg/kg) 15 min before suffered from cerebral ischemia. Neurologic function was scored at 24 h reperfusion after 2 h ischemia. HE staining was used to examine brain tissue injury, wet-dry ratio was measured to estimate cerebral edema, Western blot was used to detect the expression of p-P38 and AQP4, RT-PCR was used to detect AQP4mRNA. Results P38 inhibitor SB203580 could improve behavioral scores of ischemic reperfusion rats. The brain water content was decreased after SB203580 treatment compared with I/R group(0.792±0.010 vas 0.817 ± 0.012). The expression of p-P38, AQP4 and AQP4mRNA in margin of infarction of SB203580 group are decreased compared with I/R group compared with I/R group . Conclusion P38 inhibitor SB203580 alleviate the AQP4 expression and cerebral edema in cerebral ischemia reperfusion rats through P38 MAPK pathway.
    The effect and mechanism of phosphorus concentration on microinflammatory and oxidative stress response in maintenance hemodialysis patients
    2014, 34(3):  386-390. 
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    Objective To investigate the effect of high phosphate on mRNA and protein expression of intracellular NF-κB and NOX2/gp91 in PBMC of MHD patients and serum inflammation mediators and MDA production,so as to understand the possible relationship among hyperphosphatemia, micro-inflammatory states and oxidative stress. Method They were divided into five groups: control, MHD, dose, time and blank groups.Western blot and Real-time PCR were respectively used to determine the mRNA and protein expression of NF-κBP65、NOX2.The concentration of IL-6 in the culture supernatant was quantified by the ELISA kit and the TBA assay kit.Results NF-κB and NOX2 signal pathway in PBMCs show abnormal activation status in MHD patients. Both expression of NOX2/gp91 mRNA and protein levels increased dose-and time-dependent manners totally. IL-6 and MDA production in supernatant up-regulate dose-and time-dependent manners totally. NF-κB P65 mRNA and Phospho-NF-κBP65 protein were positively correlated with IL-6 production. NOX2/gp91 mRNA and protein expression were positively correlated with MDA production. Conclusion High phosphate could increase the production of supernatant IL-6 and MDA by activating NF-κB and NOX2 signaling pathway in PBMCs, thus could be a risk factor for microinflammatory and oxidative stress status of MHD patients.
    Silencing PARG enhance local Immune response to Colon Carcinoma
    Ge LI
    2014, 34(3):  391-396. 
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    Objective The aim of this article was to investigate the effect of silencing PARG on local immune status in colon carcinoma. Methods Lentivirus PARG-shRNA(Short hairpin RNA) was transfected into CT26 cells as experiment group, the CT26 cells without any treatment or treated with Lentivirus empty vector served as control group. Animal models for liver metastases of colon carcinoma were established by splenic subcapsular inoculation of each group of CT26 cells in Balb/c mice. The expression of PARG protein in spleen transplant tumor was detected by Western blot analysis. Immunofluorescence double labeling assay was used for the numbers detection both of B220+DEC205+DC and CD11c+CD11b+DC in the spleen. CD4+T cells and CD8+T cells in the spleen were tested by immunofluorescence single staining. All of these cells were observed by confocal laser scanning microscope. The levels of IL-10 and IL-12 in serum were measured by enzyme-linked immunosorbent assay (ELISA). Result The expression of PARG in spleen transplant tumor was reduced in PARG-silenced group compared to that in control groups(P<0.05). In PARG-silenced group, the amounts of B220+DEC205+DC were less and the numbers of CD11c+CD11b+DC were more in spleen than that in control groups(P<0.05). The numbers of CD4+T cells and the ratio of CD4+/ CD8+ in spleen of PARG-silenced group were increased compared to that in control groups(P<0.05). The serum levels of IL-10 were lower but the serum content of IL-12 were higher in PARG-silenced group than that in control groups(P<0.05). Conclusion These studies demonstrate that silencing PARG could strengthen the local immune response to colon cancer by effect on the proliferation and differentiation of DC and T cells.
    Type I antithrombin deficiency due to 10381T deletion in antithrombin gene
    2014, 34(3):  397-401. 
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    Objective We commit phenotype diagnosis and gene diagnosis aiming directly at one case of hereditary antithrombin (AT) deficiency syndrome of proband and their family phenotype, and explore the pathogenesis of family members. Methods The activity of AT(AT:A) , protein S and protein C were detected by chromogenic substrate method, with immune turbidimetry on AT antigen (AT:Ag) detection, the molecules weight and content of AT were detected by Western bloting method, Genomic DNA was extracted from blood, the 7 exons of AT and flanking sequences were amplified by PCR, products of PCR of all family members were conducted by direct sequencing analysising and gene mutation detection, screening 100 cases of normal people to exclude the of polymorphism of gene mutation. Results The AT:A and AT:Ag of proband was 48% and 121mg/L respectively, the proband′sixth exon of AT gene is c.10381T del. Some members of the family were detected the same frameshift mutations. conclusion The pedigree and some members of the first symptoms were type Ⅰ hereditary antithrombin deficiency due to AT gene 10381T del frameshift mutations.
    Progress in the study of epigenetic regulation in osteoporosis
    2014, 34(3):  406-409. 
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    Epigenetics are such heritable modifications as DNA methylation, histone modification, chromatin remodeling, as well as miRNA, without changes in DNA sequence. It has been shown that occurence of osteoporosis is closely related to the interactions between the genome and enviroment, in which epigenetic regulation play an important role.
    Advances of the CCN family in fibrosis
    2014, 34(3):  410-413. 
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    The CCN family consists of six members, namely Cyr61(CCN1), CTGF(CCN2), NOV(CCN3), WISP-1(CCN4), WISP-2(CCN5) and WISP-3(CCN6). Extensive research shows that the CCN family plays an important role in the process of fibrosis. As fibrotic inhibitors, CCN1, CCN3 and CCN5 could inhibit fibrosis; while as fibrotic promoting factors, CCN2 and CCN4 could promote fibrosis.
    Molecular Mechanism of Necroptosis and its Effect in IR injury
    2014, 34(3):  414-417. 
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    Necroptosis is a special cell necrosis that is capable of being regulated through particular molecular mechanism. Multiple stimuli could induce necroptosis , complex I, complex II and RIP1-RIP3 necrosome are critical participants in the necroptosis. Also, necrostatin-1 is a special and potent small-molecular inhibitor of necroptosis. Necroptosis could be an important alternative for cell death in ischemia-reperfusion injury.
    Advances of carfilzomib in treatment of relapsed and refractory multiple myeloma
    2014, 34(3):  418-421. 
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    Carfilzomib is a second-generation selective proteasome inhibitor that has been recently used for relapse or refractory multiple myeloma(RRMM). It has been shown to be beneficial in both bortezomib-resistant and bortezomib-naive patients, with a tolerable side effect profile. Peripheral neuropathy is less common in patients receiving carfilzomib compared to bortezomib.
    Research Progress on Bio Maker of Colorectal Cancer Stem Cell
    2014, 34(3):  422-425. 
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    CD44+、CD24+、CD166+ and CD29+ can be as colorectal cancer stem cell markers. They are involved in colorectal cancer incidence、invasion and recurrence, the higher expression of the worse prognosis. At the same time, the associated signal transduction pathways and proteases can adjust internal environment and involve in tumor formation. Based on studies of colorectal cancer stem cell surface markers, the early presence of the tumor can be detected, tumor recurrence are reduced, and find targets and breakthrough in diagnosis and treatment of colorectal cancer.
    TGF-β signaling and hematopoietic malignancies
    2014, 34(3):  426-429. 
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    Transforming growth factor β (TGF-β) is a negative regulated cytokine for growth of hematopoietic cells. TGF-β/SMAD signaling pathway takes important part in pathogenesis of hematopoietic malignance diseases. The deficiencies of TGF-β expression and mutations of TGF-β receptors or downstream signaling pathways can cause malignant proliferation of hematopoietic cells. To study the signaling pathway of TGF-β may attribute to the recognize of the pathogenesis of hematopoietic malignances and the development of therapy targeting the TGF-β pathway.
    The investigation in trainees for standardized public compulsory courses of resident training
    2014, 34(3):  430-432. 
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    Objective To evaluate the awareness of the residency policies, regulations and other related knowledge among residents attended the standardized public compulsory courses. Methods Using convenient sampling method to enroll 467 residents to fill in the questionnaire. Results Currently, the residents have an inadequate understanding about the residency policies and regulations, the correct rate is 41.2%, most of them have a better understanding in the responsibilities, obligations and rights as a doctor, the correct rate is 61%, but the awareness of law and how to protect themselves is still insufficient, the correct rate is only 47.6%. Conclusions Residents have poor understanding abort residency policies, regulations and other related knowledge, which should be improved in the future.