Table of Content

05 July 2014, Volume 34 Issue 7

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The correlation between anthropometric indexes and insulin resistance in T2DM patients after gastric bypass

2014, 34(7):  873-876. 

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Objective To discuss the correlation between anthropometric indexes and insulin resistance in T2DM patients after gastric bypass. Methods To retrospective analysis of 42 T2DM patients after laparoscopic gastric bypass, we assessed the anthropometric indexes by body fat instrument and detect islet function indicators by OGTT test before and 6 months post gastric bypass. Results After 6 months of gastric bypass, the remission rate of diabetes was 78%（32/41）. The fasting plasma glucose,body weight,body mass index,waist circumstance,hip circumstance,waist to hip ratio,body fat, insulin resistance index,insulin affective index and insulin sensitive index were significantly decreased compared with before(P<0.05). Preoperative insulin resistance index was significantly correlated with trunk fat (P <0.05). Postoperative insulin resistance was positive correlated with weight, body mass index, waist circumference, hip circumference and body fat（P<0.05）. Insulin affect index was negative correlated with body weight, body mass index, hip, hip drop value and body fat (P<0.05). Conclusion After laparoscopic gastric bypass, the changes of trunk fat content was the biggest in T2DM patients; the changes of body fat especially in abdomen in T2DM patients are associated with insulin resistance improvement.

Calcium-sensing receptor mediation of hypoxia-induced mucous hypersecretion of human airway epithelial cells by ERK signaling pathway

2014, 34(7):  877-881. 

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Objectlve To explore the signal pathway of calcium-sensing receptor (CaSR) in hypoxia-induced airway mucous hypersecretion. Methods Cultured human airway epithelial cells 16HBE by hypoxia incubator (37 oC,94%N2-1%O2-5%CO2) and divided into 6 groups:control group,hypoxia group,CaSR specific activator CaCl2 group treated with hypoxia,negative-siRNA control group treated with hypoxia,CaSR-siRNA group treated with hypoxia,extracellular signal regulated kinase(ERK) specific inhibitor U0126 group treated with hypoxia.The levels of MUC5AC mRNA were detected by RT-PCR.The protein levels of CaSR,ERK and phosphorylated ERK(p-ERK) were detected by Western blot.The levels of MUC5AC secretion were measured by ELISA.Cell survival rate was measured by MTT assay. Results CaSR protein was expressed in 16HBE and transfection with CaSR-siRNA significantly inhibited the expression of CaSR.In hypoxia group,the levels of p-ERK,MUC5AC mRNA and MUC5AC protein were increased (0.63±0.11,0.51±0.03,0.56±0.07) versus the control group (0.27±0.04,0.18±0.04,0.25±0.06) (P<0.05).CaCl2(a CaSR agonist) amplified the effect of hypoxia(P<0.05). Transfection with CaSR-siRNA or pretreatment with ERK inhibitor decreased the levels of p-ERK,MUC5AC mRNA and MUC5AC protein induced by hypoxia(P<0.05).Conclusion CaSR mediates hypoxia-induced airway mucous hypersecretion through MEK/ERK1/2 signaling pathway.

RAS signal pathway participation in interferon-α inhibition of proliferation of vascular smooth muscle cells in rats

2014, 34(7):  882-885. 

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Objective To study the action of RAS signaling pathway in interferon alpha (IFN-α) inhibition proliferation of vascular smooth muscle cells (VSMCs) in rats. Methods Cultured VSMCs were treated with transfection IFI204 siRNA and/or IFN-αin vitro instantaneously. Nonspecific siRNA transfection group was as control group. The cell vitality was detected by MTT method, and the cell cycle was analyzed by flow cytometry. The expression of P204 mRNA was determined by semiquantitative RT-PCR. P204, RAS protein and the phosphorylation levels of RAF and ERK was analyzed by Western blotting. Results Compared with control group, the expression of P204 mRNA and protein up-regulated(P＜0.01), the cell vitality and the cell cycle of G1/S transition down-regulated(P＜0.01). In the process of the above, the expression of RAS protein decreased(P＜0.01) and the phosphorylation levels of RAF and ERK droped(P＜0.01); When intervention with IFN-α after transfection IFI204 siRNA, down-regulated the expression of P204 mRNA and protein(P＜0.01), and cells of G0/G1 stage increased and S stage decreased(P＜0.01), but the expression of RAS protein and the phosphorylation levels of RAF and ERK did not increase correspondingly(P＜0.01). Conclusion RAS signal pathway participation in interferon-α inhibition of proliferation of vascular smooth muscle cells in rats.

Evaluation of proximal stomach function by simultaneous determination of intra-gastric pressure in perfusion nutrient load test

2014, 34(7):  886-890. 

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Objective To extensively evaluate the method of simultaneous determination of intra-gastric pressure (IGP) in perfusion nutrient load test (PNLT) in HS. Methods 30 HS were recruited. All HS were received simultaneous determination of IGP in PNLT. The barostat study was completed in another day. Visual analogue scale (VAS) was used to evaluate satiety during simultaneous determination of IGP in PNLT and the barostat study. Finally, all totally data were collected and analyzed. Results Pressure and volume of different satiety evaluated by VAS had significant correlation between the two methods respectively；IGP at maximal satiety was significantly correlated with peak accommodation volume and meal induced relaxation volume by the barostat study (P<0.05)；Maximal perfusion volume has significantly correlated with gastric postprandial averaged volume by the barostat study (P<0.05). Conclusions The method of simultaneous determination of IGP in PNLT has excellent feasibility, safety and reliability, and could be useful for evaluating proximal stomach function.

Molecular mechanism of C-phycocyanin in alleviation of acute lung injury in septic rats

2014, 34(7):  891-895. 

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Objective To observe the protective effect and molecular mechanism of C-phycocyanin (CPC) on acute lung injury in septic rats. Methods Rats were randomly divided into control group, model group and CPC group. Cecal ligation and puncture was used to establish a septic acute lung injury rats (model group). For the CPC groups, septic acute lung injury rats were administrated by 20,40 and 60mg/kg of CPC by peritoneal injection. 72h after the operation, serum and lung tissue were obtained, the PaO2/FiO2, the wet to dry weight ratio, content of tumor necrosis factor (TNF)-α、interleukin (IL)-6 and IL-10 in bronchoalveolar lavage fluid, the concentration of malondialdehyde (MDA) and activity of myeloperoxidase (MPO) was analyzed. Enzymic activity of heme oxygenase (HO)-1 and its mRNA was detected by colorimetric method and RT-PCR, respectively. Results The PaO2/FiO2, the content of MDA, activity of MPO, and thewet to dry weight ratio was increased in the model group, and it was statistically significant than that in the control group (P<0.05), after CPC treatment, the level of PaO2/FiO2 increased, the wet to dry weight ratio and the content of MDA and activity of MPO decreased (P<0.05), In the medel group, TNF-α、IL-6 and IL-1β in bronchoalveolar lavage fluid was increased, HO-1 acivity and mRNA level was also increased(P<0.05). CPC could decrease the cytokines level, and upregulate HO-1 acitvity and mRNA expression. In addition, HO-1 agonist CoPP could synergize CPC to production of TNF-α、IL-6 and IL-10 1β, but HO-1 antagonist ZnPP could futher increase the cytokines production. Conclusion CPC could induce HO-1 expression, and thus relieve gas exchange function and the inflammatory response in the septic acute lung injury.

The regulatory network in myeloid differentiation promoted by miR-29a and miR-142-3p

2014, 34(7):  896-903. 

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Objective To understand the regulatory network of myeloid differentiation promoted by miR-29a/miR-142-3p deeply. Methods Firstly, miR-29a and miR-142-3p were overexpressed in HL-60 cells meanwhile HL-60 cells were also induced towards monocyte and granulocyte differentiation using PMA and ATRA respectively. Used May- Grünwald-Giemsa staining to observe the nuclear morphometry change and utilized Real time PCR to detect the expression of CD11 and CD14 in cells. Genes which possessed similar expression pattern were obtained by microarray and analyzed for their similarities. Analyzed by GO and signal pathway clustering using DAVID and GeneMANIA. Utilizing PicTar and TargetScan, predicted new targets of miR-29a which were also verified by double-luciferase reporter assay. Results Many differential expressed genes during ATRA/PMA induced myeloid differentiation or in the cells with miR-29a/miR-142-3p overexpression were similar. Significantly enriched genes in GO analysis which also possessed similar expression patterns were almost associated with cell differentiation. Conclusion miR-29a and miR-142-3p regulated the activity of their targets, triggered global gene expression change to make it closer to that during myeloid differentiation and then promoted myeloid differentiation. ZBTB5, CaMKK2, SUV420H2, TRIB2 and CANX may play important roles in myeloid differentiation promoted by miR-29a.

The Effect of Humanized CDR3δ1-Grafted Antibody on PBMC-Mediated Treatment of Human Liver Cancer

2014, 34(7):  904-908. 

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Objective To investigate the effect of humanized CDR3δ1-grafted antibody GTM-Fc on PBMC-mediated treatment of human hepatoma. Methods HepG2.2.15 were transplanted by subcutaneous injection to form transplantation tumor in nude mice. Then were divided into PBS group, PBMC group and PBMC+Ab group, nine mice in each group. Mice in PBS group were injected within tumor with 100μL PBS(phosphate buffer solution) while those in PBMC group and PBMC+Ab group were injected respectively with 1×106 of PBMC and 1×106 of PBMC added 3ug GTM-Fc. Repeated the treatment every three days and measured the weight and the tumor size. executed all the mice three day after the fith treatment. Flow cytometry(FCM) was performed to detect the binding capability of GTM-Fc to human hepatic carcinoma cells(HHCC). The antibody-dependent cell-mediated cytotoxicity (ADCC) was applied to detect the effect of GTM-Fc on PBMC-mediated killing HHCC in vivo and in vitro. Results Humanized CDR3δ1-grafted antibody GTM-Fc displayed excellent binding activity to HHCC. Experiments in vitro showed that GTM-Fc augmented the killing effect of PBMC to HHCC. Conclusions Humanized CDR3δ1-grafted antibody GTM-Fc can enhance the killing effect of PBMC on HHCC.

HMGB1-TLR4 promote anoxia/reoxygenation injury-mediated apoptosis in rat proximal tubular epithelial cells

2014, 34(7):  909-913. 

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Objective To study the expression of high mobility group box-1 protein (HMGB1) and Toll-like receptor 4 (TLR4) in renal proximal epithelial cells following by anoxia/reoxygenation injury and their underlying roles in cell apoptosis. Methods Rat primary proximal tubule epithelial cells (PTECs) were cultured in six-well plates and randomly divided into Control, anoxia/reoxygenation group(A/R), HMGB1 antibody treatment group (A/R+HMGB1) and TLR4 antibody treatment group (A/R+TLR4). RT-PCR was used to detect the mRNA levels of HMGB1 and TLR4; Western blot was used to detect the protein expression of HMGB1, TLR4 and the apoptosis-related factors, such as Bcl-2, Bax，CHOP and caspase-8; Flow cytometry was used to detect apoptosis. Results The expression of mRNA and protein of TLR4 and HMGB1 increased significantly in A/R group compared with control (P<0.01). The apoptosis cells in A/R group increased significantly compared with control (P<0.01). The expression of Bax, CHOP and caspase-8 increased significantly compared with control (P<0.01). While the expression of Bcl-2 decreased compared with control (P<0.05). After HMGB1 and TLR4 antibodies treatment, the apoptosis cells in A/R+HMGB1 group and A/R+TLR4 group decreased significantly compared to A/R group (P<0.05). The expression of Bax, CHOP and caspase-8 also decreased significantly compared with A/R group (P<0.05). The change of Bcl-2 protein expression was not obviously. Conclusions HMGB1-TLR4 axis promote apoptosis of PTECs following A/R injury.

Peripheral blood monocyte hepcidin in patients of non-Hodgkin's lymphoma may lead to anemia of chronic disease

2014, 34(7):  914-920. 

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Objective The role of monocytes hepcidin in NHL patients.Methods Collecting the clinical information and peripheral venous blood of non-Hodgkin’s lymphoma patients.Serum concentration of IL-6 and TNF-α was detected by ELISA. Peripheral blood monocytes were isolated by CD14+ Magnetic beads.Hepcidin, IL-6 and TNF-α mRNA of monocytes were detected by Real time PCR quantitative assay.Results In NHL patients, hemoglobin was reduced(112.5±21.7g/l),monocyte hepcidin expression was higher than normal（P=0.013）.Monocyte hepcidin in untreated patients was negatively correlated with Hb and IL-6 levels(P<0.05),but not associated with TSAT%, SF and TNF-α levels.Conclusions The increased monocyte hepcidin in untreated lymphoma patients may play an etiologic role in ACD.

Thymosinβ4 improves CD31 and eNOS expression in rat cerebral cortex after focal cerebral ischemia/reperfusion by up-regulation phosphorylation level of Akt

2014, 34(7):  921-926. 

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Objective To investigate the effect and mechanism of Thymosin beta4(Tβ4) on angiogenesis in rat cerebral cortex after focal cerebral ischemia/reperfusion. Methods Rats were randomly divided into sham group(S), model group(IR), Tβ4 group(IRT), Tβ4+LY294002 group(IRTL), 0.9% sodium chloride injection group(IRN). The focal ischemia/reperfusion rat model was established by filament occluding the right middle cerebral artery for 2 hours. Rats were divided into 3 and 7 day subgroups in IR、IRN and IRT group. The protein expression of eNOS、p-Akt and Akt were detected by western blotting. Immunohistochemical method was selected to detect the expression of Tβ4 and CD31. The mRNA expression of eNOS、SDF-1 and CXCR4 were tested by fluorogenic quantitative PCR. Longa score was selected to evaluate neurobehavioral. Results Compare with IRN group, the protein expression of Tβ4 increased significantly at IRT 3 day(P<0.01); compared with IRN group, the protein expression of eNOS、p-Akt increased significantly at IRT 3 and 7 day (p<0.05), but in IRTL group they reduced significantly (P<0.01); compare with IRN group , the mRNA expression of eNOS、SDF-1 and CXCR4 were increased significantly at IRT 3 and 7 day (p<0.05); The number of CD31 postive micrangium increased significantly at IRT 3 and 7 day (P<0.05) and the neural function recovery significantly at IRT 7 day (p<0.05). Conclusion Tβ4 may promotes angiogenesis and behavioral recovery by increasing p-Akt and eNOS expression in a rat model of focal cerebral ischemia/reperfusion.

Phenethyl isothiocyanate inhibits human colon cancer SW480 cells migration and invasion by PI3K-NF-κB-MMP-9 pathway

2014, 34(7):  927-932. 

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Objective To investigated the anti-invasive mechanisms of PEITC in human colon cancer SW480 cells. Methods Human colon cancer SW480 cell line was cultured in vitro, and treated with 10, 30,50μmol/L PEITC for 24h. Cell viability, cell migration and invasion was analyzed by MTT assay, scratch healing assay and transwell membrane assay, respectably. Activity of Matrix metalloproteinase-9 (MMP-9) was analyzed by a fluorescence resonance energy transfer kits, and MMP-9 mRNA expression was detected by RT-PCR. Activation of phosphoinisitide-3-kinase (PI3K) /Akt/mammalian target of rapamycin (mTOR) and phosphatase and tensin homologue (PTEN), and nuclear translocation of nuclear factor kappa B (NF-κB) were examined by Western blot. NF-κB activity was detected by luciferase reporter assay. Furthermore, a mice model with xenograft tumors was constructed, and the effect of PEITC on tumor growth was analyzed. Results PEITC significantly reduced SW480 cells invasion and migration without affecting the viability of cells (P<0.05). PEITC also markedly inhibited MMP-9 activity and expression at both protein and mRNA levels (P<0.05). PEITC attenuated PI3K and phosphorylation of AKT and mTOR, whereas PTEN was increased. In the meantime, PEITC inhibited NF-κB transcriptional activity (P<0.05). In addition, PEITC diminished NF-κB nuclear translocation. Furthermore, PEITC could significantly inhibit xenograft tumors growth in nude mice (P<0.05). Conclusion PEITC as a potential anti-invasive agent by inhibiting MMP-9 involved in PI3K/AKT and NF-κB pathways.

Effects of acute low dose radiation on DNA methylation in liver and bone marrow of mice

2014, 34(7):  933-938. 

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Objective To study the effect of acute low dose radiation on DNA methylation and DNA methyltransferase(DNMT) in liver and bone marrow of mice . Methods The C57BL/6 mice was irradiated by linear accelerator with X ray one time. the peripheral blood cells were made in 1、3、7、14 days after radiation, Then these mice were killed , liver and BMNC was exacted. The DNA methylation levels of whole gene were detected by HPLC, the expression of DNMT1、DNMT3A and DNMT3B were detected by Western blot(WB). Results Compared with those in the normal group, the numbers of peripheral leukocyte and platelet was decreased significantly in the radiation group, the peripheral leukocyte in the first day dropped to the lowest and platelet fell to lowest in the third day, they all were not returned to normal in the 14rd. the DNA methylation levels of radiation group was decreased significantly, but with the increase of time, the methylation level returned to normal. the expression of DNMT1 and DNMT3A were increased significantly in the radiation group, they all were not returned to normal in the 14rd,the expression of DNMT3B were increased significantly, but returned to normal in 7rd day. Conclusion Acute low dose radiation can decrease the methylation levels of DNA and the expression of DNMTs.

Analysis of difference in histone H3K9 trimethylations between heart and spleen of the normal human Organs by ChIP-seq

2014, 34(7):  939-944. 

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Objective In this work, it showed that modifications are closely-associated with tissue-specific expression, function and development by generating the genome-wide maps of H3K9me3 of human heart and spleen. Methods The DNA of samples were extracted by Chromatin immunoprecipitation. The ChIP results were validated by qPCR.Then building the ChIP Sequencing library, contrasting with the target genome sequence to obtain compared Reads. And peak analysis the whole genome, GO enrichment analysis the biological functions of the related genes of peak. Results It found that 169 genes displayed significant H3K9me3 differences between heart and spleen. Among these genes, 64 genes are the special genes in heart-H3K9me3; 87 genes are the special genes in spleen-H3K9me3. From these genes, select eight genes of H3K9 which were significantly expression. Then selecting two genes of H3K9 which were the most significant difference expression from the eight genes.They are PTPN3 and RBMS. Conclusion Above the findings show the significance of H3K9me3 as a potential biomarker or promising target for epigenetic-based disease treatment.

Celecoxib induces intervertebral disc cells expression of nerve growth factor

2014, 34(7):  945-949. 

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Objective To investigate the effect of a selective COX-2 inhibitor Celecoxib and PGE2 on expression of nerve growth factor (NGF). Methods Isolated human IVD cells were stimulated with 10ng/ml of IL-1β in the presence or absence of different concentration of Celecoxib. NGF expression and paoduction were determined by real-time PCR and ELISA, respectively. Secretion of PGE2 was determined by ELISA. The effects of exogenous PGE2 and its receptor (EPs1-4) agonists were also tested for NGF production. Results Stimulation of IL-1β for 3h could induce IVD cells to express NGF mRNA in a dose-dependent manner. Pre-treatment with Celecoxib strongly enhanced it to 1.8 fold. 10ng/mL of IL-1β induced PGE2 production to (5.46±0.64)ng/mL, At the concentration of 10mmol/L Celecoxib, PGE2 release was substantially inhibited to (1.07±0.04) ng/mL (P<0.05). 100μmol/L PGE2 inhibited IL-1β induction of NGF to 59% (P<0.05), and this effect was mimicked when agonists EP2 and EP4, but not EP1 and EP3, were supplemented to the culture. Conclusion. Celecoxib can enhance IL-1β-induced NGF expression, and this effect may be mediated by inhibition of PGE2 production.

Dexamethasone promotes apoptosis of primary human thyroid follicular epithelial cells

2014, 34(7):  950-954. 

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Objective To investigate the effect of dexamethasone on apoptosis of primary human thyroid follicular epithelial cells. Methods After separation and stimulation with different concentrations of dexamethasone, inhibition rates of proliferation, apoptosis rates and apoptosis-related mRNA expression of primary thyroid follicular epithelial cells were measured by MTT, flow cytometry and fluorescence quantitative PCR respectively. Results Within the concentration range of 10-6~10-4mol/L, dexamethasone dose-dependently inhibited thyroid cell proliferation, increased cell apoptosis rates and expression of apoptosis-related mRNA of thyroid cells. Conclusion Dexamethasone can not only inhibit the proliferation of thyroid follicular epithelial cells, but also have effect on apoptosis of thyroid cells.

Construction of luciferase reporter gene vectors containing B7-H1 promoter region and promoter activity assay

2014, 34(7):  955-960. 

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Objective To construct luciferase reporter gene vectors containing B7-H1 promoter region，and detect promoter activity in HepG2 cells. Methods Firstly several fragments of the B7-H1 gene 5’-UTR promoters were amplified by PCR and cloned into pGL3 basic vector. After the identification by digestion and sequencing on the recombinant basic vector, they were named p88，p175, p203, p398, p723, and p960 respectively. Then one of the human HCC cell lines, HepG2 cells, was transfected with these constructed plasmids. The expression of luciferase before and after the addition of IFN-γ was detected by Dual-Luciferase(LUC) Reporter Assay System kit. Results Five luciferase reporter gene vectors containing B7-H1 promoter region were constructed successfully identified by restriction endonuclease analysis and sequences analysis. The recombinant constructs were transiently transfected into HepG2 cells. After the addition of IFN-γ, the results of LUC detection assay showed that the expression of LUC was significantly enhanced by P175, P203, P398, P723 or P960，but not influenced by P88. Conclusion Luciferase reporter gene vectors containing B7-H1 promoter region were constructed successfully，which may be valuable for further study on B7-H1 promoter activity and molecular mechanism.

Establishment leukemia cell lines expressing IL-21 and analysis of anti-tumor function

2014, 34(7):  961-967. 

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Objective To establish leukemia cell lines expressing IL-21 constitutively, and analyze the activation function on lymphocytes by those cell lines. Methods Four leukemia cell lines were infected with lentivirus carrying IL-21 and RFP genes. Fluorescent microscope was used for observing the expression of RFP and flow cytometry was used for analyzing the percentage of RFP-positive cells. The concentration of IL-21 in the medium was detected by ELISA. After stimulation with the IL-21 stable-expressing leukemia cell lines, the amplification folds of PBMC were calculated by cell counter and the surface markers were analyzed by flow cytometry, while their cytotocicity towards different tumor cells were assessed with CCK8 test. Results Four leukemia cell lines stable-expressing IL-21 were established, namely, K562-IL-21, THP-1-IL-21, Jurkat E6-1-IL-21 and RPMI8226-IL-21. The lymphocytes amplification of 3 different samples varied from 1.147±0.057 to 2.725±0.345 times and the amplification folds of lymphocytes stimulated by different numbers of leukemia cells varied from 1.127±0.152 to 2.213±0.200. The percentage of CD3+T cell and CD56+ NK cell decreased（P<0.05）while CD19+ B cells increased（P<0.05） after stimulation. The killing rates of activated lymphocytes are much higher than the control. The activated lymphocytes had powerful killing capacity on their stimulating leukemia cells and 7 other tumor cells and the killing rates ranged from （28.68±9.31）% to （78.45±0.61）%. Conclusions Four leukemia cell lines stable-expressing IL-21 could activate PBMC to kill different tumor cells.

Rosiglitazone ameliorates intestinal barrier dysfunction induced by bile duct obstruction in mice?

2014, 34(7):  968-973. 

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Objective: To investigate the alteration of bile duct ligation (BDL) induced intestinal barrier dysfunction in mice and the protective effect of PPARγ pathway activation. Methods: The mice were divided into five groups randomly: Sham group, Sham+RGZ group, BDL group, BDL+RGZ group and BDL+RGZ+GW9662 group. All BDL group were established the model of bile duct ligation induced intestinal barrier dysfunction in mice. Mice were treated with RGZ, GW9662 or vehicle for 2 days before surgery and 5 days after. Blood and tissue samples were collected at the end of day 5. Intestinal morphological alteration were assessed, PPARγ associated oxidative stress and apoptosis and systematic inflammations of each group were determined by biochemical analysis; Protein expressions of PPARγ, Occludin, Bcl-2 and Bcl-xL in each group were evaluated by western blot. Results: BDL resulted in significant damage in intestinal barrier and severe systematic inflammation, accompanied by the protein expression of PPARγ, Occludin, Bcl-2 and Bcl-xL suppression (P<0.05); Pretreatment of RGZ significantly improved BDL induced intestinal injury and systematic inflammation, and up-regulated the protein expression of PPARγ, Occludin, Bcl-2 and Bcl-xL in the intestine(P<0.05). However, such protective effects were ameliorated in the BDL+RGZ+GW9662 group(P<0.05). Conclusion: PPARγ activation by rosiglitazone plays a protective role in intestinal barrier dysfunction induced by BDL in mice. This protective effect may be attributed to the anti-oxidant and anti-apoptosis properties of PPARγ.

Effect of siRNA-mediated silencing of the decay accelerating factor(CD55) on biological behaviour of human pancreatic cancer cell line BxPC-3.

2014, 34(7):  974-978. 

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To study the specific effect of silencing of CD55 gene by small interfering RNA (siRNA) on the growth of human pancreatic cancer cell line BxPC-3. Methods Three CD55-specific siRNAs and one negative siRNA were designed and was transfected into human pancreatic cancer cell line BxPC-3 by Lipofectamine 2000.Quantitative real-time PCR(qRT-PCR) and Western blot were used to test the expression levels of CD55 mRNA and protein respectively. And flow cytometry were used to detect apoptosis, proliferation and cell cycle of target cells. Results: One of three CD55-specific siRNAs, siRNA3 could silence the expression of the CD55 gene. The expression levels of CD55 mRNA and protein decreased remarkably.The cells transfected with siRNA3 were found to increase apoptosis and reduce proliferation significantly;the target cells stagnated in phase G1 after transfection(P<0.05). Conclusion: SiRNA-mediated silencing of the decay accelerating factor(CD55) can effectively affect the biological behavior of human pancreatic cancer cell line BxPC-3.

Role of silent information regulator 1（SIRT1） in the malignant biological behavior of prostate cancer

2014, 34(7):  979-983. 

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Objective　To detect the expression of silent information regulator 1（SIRT1）in the clinical prostate cancer tissues and cells, and to explore the role of SIRT1 in the development of prostate cancer. Methods A total of 8 human prostate adenocarcinoma tissues and adjacent normal prostate tissues were acquired from the First Affiliated Hospital of Liaoning Medical University and the SIRT1 and P53 mRNA and protein expression levels in prostate tissues were determined by Real-time PCR and Western blot. The normal human prostate epithelial cells PrEC and human prostate carcinoma cell lines LNcap, PC3and DU145 were cultured in vitro, then the SIRT1 and P53 mRNA expression levels were detected by real-time PCR. The whole cell protein lysates, nuclear and cytosolic protein lysates were prepared for Western blot to determine the SIRT1 and P53 protein expression levels in PC3 and DU145 cells. Results SIRT1 was found to be overexpressed in human prostate cancer tissues compared with adjacent normal prostate tissue（P<0.01）. SIRT1 was significantly overexpressed in human prostate cancer cells LNcap, PC3 and DU145 compared with PrEC cells at mRNA and protein levels （P<0.01）and SIRT1 was predominantly localized in the nuclear of PC3 and DU145 cells（P<0.01）. Conclusion Nuclear SIRT1, via P53 dependent and independent activation, could contribute to the development of prostate cancer, and pay a foundation for exploring the possible role and underlying mechanisms of SIRT1 on the malignant behavior of prostate cancer.

Increased bile cholesterol content inhibits the gallbladder contraction by affecting the re-distribution of CCK-1R

2014, 34(7):  984-989. 

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Objective To analyze the relationship between bile total cholesterol (TC) content and the contraction of the gallbladder, and to explore its possible mechanism. Methods A total of 73 patients with or without gallstones, who underwent biliary surgery in the Department of General Surgery in PUMCH from March 2013 to January 2014, were selected and divided into gallstone group and non-gallstone group. The bile of all these patients was collected during the operation, and the content of the bile TC and total bile acid (TBA) was detected. The gallbladder contraction test in patients with gallstones was performed before the surgery. The expression of cholecystokinin-1 receptor (CCK-1R) and caveolin-3 (Cav-3) in gallbladder mucosa and the smooth muscle in two groups was detected using immunohistochemistry staining method. The co-localization of CCK-1R and Cav-3 in gallbladder mucosa and the smooth muscle in two groups was also observed using immunofluorescence method. Results The content of bile TC and TBA in patients with gallstones was significantly higher than that in patients without gallstones (P<0.05). And the bile TC content in patients with a worse contraction of the gallbladder was significantly higher than that in patients with a better contraction of the gallbladder (P<0.05). The immunohistochemistry staining revealed that the expression of Cav-3 in the gallbladder mucosa in gallstone group was significantly higher than that in non-gallstone group (P<0.05). The co-localization of CCK-1R and Cav-3 in gallstone groups was obvious in both gallbladder mucosa and the smooth muscle, while no obvious co-localization was observed in non-gallstone group. Conclusion The increased bile TC content can inhibit the contraction of the gallbladder, and the mechanism may relate to excess Cav-3 binding with CCK-1R, thus affecting the function of CCK.

Analgesia Nociception Index is a new monitoring index for intraoperative analgesia evaluation in inhalation anesthesia

2014, 34(7):  990-993. 

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Objective To evaluate the performance of analgesia nociception index (ANI) measured during general anesthesia with inhalational anesthetics for laparoscopic cholecystectomy. Methods 32 ASAⅠ~Ⅱ patients undergoing laparoscopic cholecystectomy with general anesthesia using sevoflurane were included in this observational study. The depth of anesthesia was maintained from D0 to D2 by Narcotrend (NT) monitoring. The ANI (100 maximum optimal value, 0 minimal value) as well as mean blood pressure (MAP) and heart rate (HR) were observed at 5 different time point: no nociceptive stimuli after induction (T0), laparoscopic pneumoperitoneum (T1), trocar puncture into the abdomen (T2), gallbladder bed dissection (T3), gallbladder removal from abdomen (T4). Results With the same anesthesia depth, ANI decreased significantly (P<0.05) in nociception point T1, T2, T3 and T4, compaired with no stimuli point (T0). The most intense painful stimulation is trocar puncture into the abdomen (T2) of which ANI decreased most rapidly and significantly (40.0±11.4, p<0.001). Conclusions ANI appears to reflect different levels of stimulation during sevoflurane based general anesthesia.

Analyzing about Genetic Traits of Hair and Hand among Han nationality in North Guizhou

2014, 34(7):  994-995. 

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The relationship between STAT3 pathway and paclitaxel-resistance in breast cancer

2014, 34(7):  996-997. 

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Research advances in peptide drugs and anti-tumor

2014, 34(7):  998-1001. 

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In the anti-tumor treatment, anti-tumor peptide drug has unique advantage: low molecular weight, fewer adverse reactions, anti-tumor specificity, low immunogenicity, and ease of synthesis and transformation. It will be great value to clinical treatment.

The research advances in the role of telomere/telomerase in premature ovarian failure

Wei-Yu WANG

2014, 34(7):  1002-1005. 

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Ovarian function has close association with division, proliferation and differentiation of oocytes and granulosa cells in ovary. Telomere, as a molecular clock measuring the ability of cell division, plays a critical role in division and differentiation of variant eukaryotic cells. Recent researches summerized premature ovarian failure based on telomere and telomerase.

Research progresses of cancer stem cells as Emerging targets in pancreatic cancer

2014, 34(7):  1006-1009. 

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Pancreatic ductal adenocarcinoma is one of the most aggressive malignancies and is characterized by poor response to current therapy and a dismal survival rate. Recent insights regarding the role of cancer stem cells (CSC) in tumorigenesis have brought further understanding of the field and have highlighted new therapeutic targets. CSCs are a distinct subset of cancer cells, with the ability to differentiate into other cell types and self-renew in order to fuel the maintenance of tumor amplification. Here review the key pathways involved in these processes, the biomarkers used to identify CSC for finding new therapeutic approaches targeting CSC.

Recent advances of Sirtuins in chronic obstructive pulmonary disease

2014, 34(7):  1010-1013. 

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Sirtuins are class III histone deacetylase. SIRT1 and SIRT6 are the major members of Sirtuins participated in the development of chronic obstructive pulmonary disease(COPD). The expression of SIRT1 and SIRT6 were down-regulated in the lungs of patients with COPD, and they regulate inflammatory reaction, lung autophagy and senescence through multiple mechanisms. Sirtuins may be one of main regulatory molecules family in the progression of COPD.

Impact of a web-based study of genetic disorders on genetics education in the context of genomic medicine

2014, 34(7):  1014-1016. 

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Objective To summarize the impact of a web-based study of genetic disorders in the 8-year program medical students at PUMC in the post-genome era. Method A survey for the impact of this practice on genetic education was designed and collected from students. Result This practice is helpful for students to understand “genomic medicine”, to expand their scientific perspective and to stimulate their interests to scientific research, to some extent. Conclusion The integrated case-based learning would be the trend for the genetics education in the context of genomic medicine.