Table of Content

05 March 2016, Volume 36 Issue 3

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Effect of miR-126 knock down on the development of mouse thymocytes

2016, 36(3):  289-294. 

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Objective: To investigate the influence of miR-126 knock down on the development of mouse thymocytes and explore its significance preliminarily. Methods: The volume, weight and total cells number of thymus in miR-126 knockdown (KD) mice were conventional detected. The expression of miRNA-126 in thymus also was detected by Real-time PCR. The pathologic morphology change of thymus was observed by HE staining. The proportion, as well as the expression of nuclear antigen Ki-67 and the apoptosis of lymphocytes in thymus from miR-126KD mice were determined by Flow cytometry analysis. Finally, the expression level of p-AKT and p-ERK1/2 in the thymus were detected by Western blot. Results: Compared with those of WT mice, the volume, weight and total cells number of miR-126KD mice thymus were no obvious differenced. The expression level of miRNA-126 in miR-126KD mouse thymus was significantly lower than that in the WT mice(P<0.05). Thymus morphology of miR-126KD mice was abnormal. Compared with WT mice, the percentage and absolute number of thymic CD4+ SP cells in miR-126KD mouse were increased obviously (P<0.05), however, the proportion and number of DP cells were significantly decreased(P<0.05). Furthermore, compared with that in WT mice, the expression of nuclear antigen Ki-67 in thymic CD4+ SP cells from miR-126KD mouse were abnormally increased and the apoptosis of thymic CD4+ SP cells were decreased(P<0.05); Conversely, the expression of nuclear antigen Ki-67 in DP cells were significantly decreased(P<0.05). Finally, the expression level of p-AKT and p-ERK1/2 in the thymus were decreased in miR-126 KD mice (P<0.05). Conclusion: miR-126 knock down could affect the development of thymocytes in the thymus, especially on the development of CD4+T cells, which provide an important experimental basis for further exploring the roles of miR-126 in the development and function of thymocytes.

The autophagy induced by IFN-γ inhibit the proliferation of human placental mesenchymal stem cells of fetal side

2016, 36(3):  295-300. 

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Objective To investigate the activation of autophagy induced by IFN-γ in fPMSCs cultured in serum-free medium, so as to determine the role of autophagy in the capacity of proliferation in MSCs. Methods Enzyme dissociation was used in the isolation of MSCs of fetal side from human placenta. The characteristic of MSCs were identified by flow cytometry analysis and differentiation culture system. After fPMSCs were treated with 50?g/L IFN-γ, the expression of autophagy marker gene LC3Ⅰ/Ⅱ was measured by western blot assay; fPMSCs were infected with mRFP-GFP-LC3 adenovirus, the puncta light was observed by confocal fluorescence microscopy; MTT assay was employed to identify the effect of autophagy induced by IFN-γ in fPMSCs for the capacity of proliferation. The normal cells and the cells treated with 3-Ma was as control sets in the same way. Results The cells isolated from human placenta of fetal side expressed MSCs surface marker CD73, CD90 and CD105, but did not express CD14, CD34 and CD45, and had the ability of differentiation into adipogenic and osteogenic cells. IFN-γ could increase the transfer ratio of LC3Ⅰ to LC3 II (P <0.05), and induce more puncta light existed in cytoplasm by Confocal fluorescence microscope. 3-Ma could alleviate the inhibition of proliferation carried by IFN-γ in fPMSCs (P <0.05). Conlusion The autophagy induced by IFN-γ negatively regulated the ability of proliferation in fPMSCs.

Establishment and identification of OVCAR3 cell line stably expressing inducible shRNA targeting CKS2

2016, 36(3):  301-306. 

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Objective To establish a OVCAR3 cell line stably expressing inducible shRNA targeting CKS2 and explore the effects of CKS2 knockdown on cell proliferation of OVCAR3. Methods Two oligonucleotides targeting CKS2 gene were synthesized and cloned into lentivirus expression plasmid pLVTHM, thus, pLVTHM/CKS2 shRNA recombinant plasmids were constructed. OVCAR3 cells were firstly infected with lentivirus made from pLV-tTR/KRAB-Red empty vector, followed by infecting with another kind of lentivirus prepared from pLVTHM/CKS2 shRNA recombinant plasmid. The expression of CKS2 mRNA and protein were determined by real time PCR and Western blot in OVCAR3 cells infected with lentivirus and treated with doxycycline (DOX) for 72h, respectively. The effects of cell proliferation were determined by MTT. Results The lentivirus expression plasmids containing CKS2 shRNA were constructed successfully, the lentivirus were produced and OVCAR3 cells were infected by these lentivirus. A OVCAR3 cell line stably expressing inducible shRNA targeting CKS2 was established successfully and the CKS2 expression levels were decreased obviously after induction by DOX（P < 0.05，P < 0.01）. CKS2 knockdown could inhibit OVCAR3 cell proliferation. Conclusions A OVCAR3 cell line stably expressing inducible shRNA targeting CKS2 was established successfully, CKS2 expression suppression induced by DOX could inhibit OVCAR3 cell proliferation.

Effect of 5-Aza-CdR on miR-1247-5p expression and its gene methylation degree in promoter region in hepatocellular carcinoma cell line SMMC-7721

2016, 36(3):  307-310. 

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Objective To investigate the different expression of miR-1247-5p and its CpG island methylation on gene promoter region between normal human liver cell line L-O2 and hepatocellular carcinoma cell line SMMC-7721 by the intervention of different concentration of 5-Aza-CdR. Methods Normal human liver cell line L-O2 and hepatocellular carcinoma cell line SMMC-7721 were treated with different concentrations of 5-Aza-CdR(0, 5 and 10 μmol/L), and then to exam the methylation of miR-1247-5p gene promoter region of SMMC-7721 cell line by methylation-specific PCR (methylation specific polymerase chain reaction, MSP), finally, to detect the expression of miR-1247-5p by SYBR Green qReal Time PCR. Results Compared with normal human liver cell line L-O2, the expression of miR-1247-5p in hepatic carcinoma cell line SMMC-7721 was decreased（P < 0.05） and the methylation of its gene CpG Island in promoter region was higher. The expression of miR-1247-5p was significantly raised compared with the control group (p < 0.01) and the methylation of miR-1247-5p gene CpG Island in promoter region was reduced after demethylation drug intervention. Conclusion The methylation of miR-1247-5p expression might be involved in the occurrence and development of hepatocellular carcinoma．

BEZ235 inhibits the proliferation of breast cancer cells in vitro and in vivo

Song-Nan ZHANG Zhen-Hua Lin Tie-Feng Jin

2016, 36(3):  311-314. 

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Objective To investigate the effect of BEZ235 on breast cancer cells. Methods Proliferation ability of human triple negative breast cancer cell line MDA-MB-231 and triple positive breast cancer cell line MCF-7 was detected by MTT assay; the xenograft model to analysis the ability of exhibited breast cancer proliferation in vivo; Western blot method to detect the PI3K/Akt signaling pathway related protein level. Results BEZ235 significantly inhibited the proliferation of breast cancer cells (p<0.01). Especially BEZ235 can strongly inhibited the triple negative breast cancer MDA-MB-231 cells in vivo and in vitro (p<0.01). Conclusion BEZ235 may through the inhibition of PI3K/Akt signaling pathway proteins phosphorylation to inhibition of breast cancer cells in vitro and in vivo.

Interaction between TGF-βRI and Fascin1

2016, 36(3):  315-320. 

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Objective To probe the interaction between TGF-βRI and Fascin1. Methods The co-localizaion of TGF-βRI and Fascin1 was verified by immunofluorescence. Meanwhile, GST pull-down and Co-IP assay were used to identify the interaction and the binding site between TGF-βRI and Fascin1. Results The immunofluorescence experiment showed that the two proteins localized at the same places. TGF-βRI was accociated with Fascin1. And the N-terminal of Fasicn1 and ICD domain of TGF-βRI were invovled in the interacton. Conclusion TGF-βRI interacts with Fascin1, and the N-terminal of Fasicn1 and ICD domain of TGF-βRI were invovled in the interacton.

Iron overload injures Sertoli cells of mouse

2016, 36(3):  321-326. 

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Objective To investigate the effect of iron overload on sertoli cells. Methods sertoli cells were isolated and cultured by adding iron dextran at different concentrations (50,100,200μmol/L ) for different lengths of time (24,48,72h). Iron overload was observed by microscopy and flow cytometry. ROS production was observed by flow cytometry. GSH and GSH-Px contents were determined by nitrobenzoic acid method.MDA content was measured by thiobarbituric acid way.NO and NOS contents were detected by nitrate reductase way.SOD content was examined by xanthine oxidase method.The expression levels of occluding,ABP,TRF,INH and VIM protein were observed by Western blot and immunocytochemistry. Results Compared with control group, the level of intracellular ROS, the contents of MDA, NO and NOS all significantly increased, with consequent decrease in the contents of SOD, GSH, GSH-Px (P＜0.01).The expressions of occluding,ABP,TRF,INH and VIM were also significantly decreased (P＜0.05).Conclusion Iron overload can induce oxidative damage of sertoli cells and weaken its function.

Effect of miR-147 on the production of proinflammatory cytokines in macrophages from aging mice

2016, 36(3):  327-330. 

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Objective To estimate the effect of miR-147 on innate immune response of macrophages from aging mice. Methods The levels of IL-6 and TNF-α were detected by ELISA in serum of young and aging mice injected intraperitoneally with LPS. The expression of miR-147 was detected by real-time quantitive PCR (q-PCR) in LPS-treated peritoneal macrophage of mice. The level of IL-6 and TNF-α was detected in mouse peritoneal macrophages transfecetd with mimics or inhibitor of miR-147. Results The level of proinflammatory cytokine IL-6 and TNF-α were higher in the aging mice than that in the young group (P< 0.05). The level of miR-147 increased significantly in mouse peritoneal macrophages from the young mice compared to the aging mice (P<0.05). Overexpression or down-regulation of miR-147 decreased or increased the level of IL-6 and TNF-α dramatically in LPS-stimulated mouse peritoneal macrophages (P<0.05). Conclusions The change of miR-147 level in peritoneal macrophages from aging mice may be an important mechanisms leading to disorder of proinflammatory cytokine production in aging mice.

Inhibition of nitric oxide on the activation of NLRP3 inflammasome myoblast /myotubes stimulated by IFN-γ in vitro

2016, 36(3):  331-336. 

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Objective: To observe the effect of sodium nitroprusside (SNP) or nitro L arginine acid methyl ester (L-NAME) on the NLRP3 inflammasome activation of C2C12 myoblast or differentiated myotubes in inflammatory culture in vitro. Methods: C2C12 myoblasts were stimulated by IFN-γ, then the formation and activation of NLRP3 inflammasome were analyzed by qPCR and Western blot,and the secretion of IL-1β in the cell culture supernatant was detected by ELISA. Further, C2C12 myoblasts induced by IFN-γ were treated by L-NAME or SNP respectively, then the formation and activation of NLRP3 inflammasome and the secretion of IL-1β were analyzed. Results: qPCR and Western blotting detection confirmed that expression of NLRP3, ASC and mature-caspase-1 in C2C12 myoblasts or differentiation myotubes induced by IFN-γ was up-regulated（P<0.01）. ELISA analysis further confirmed that the IL-1β concentration in the medium of C2C12 induced by IFN-γ was up-regulated compared with that of the non stimulating cells（P<0.01）.At 6 h after SNP treatment, a significant down-regulation of mRNA and protein levels of differentiation myotube inflammasome (ASC and NLRP3, caspase-1) was found（P<0.01） and a contrary result was obtained after L-NAME treatment（P<0.05）. Consisvent with the above results, the expression of IL-1βin the culture supernantent was down-regulated or up-regulated by SNP or L-NAME respectively（P<0.05）. Conclusions: In inflammatory conditions, C2C12 myoblasts or differentiation myotubes possess the ability to synthesize inflammasome NLRP3. NO might cause a certain inhibition on the NLRP3 inflammasome formation and activation.

Somatostatin receptor subtype-4 agonist mitigates isoflurane induced cognitive dysfunction in aged rats

2016, 36(3):  337-341. 

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Object To investigate the effect of somatostatin receptor subtype-4 agonist(NNC 26-9100) on learning and memory impairment in aged rats induced by isoflurane anesthesia. Methods 20-month-old Sprague Dawley rats were randomly divided into 4 groups(n=14) as follows: V+O2, V+ISO, NNC+ISO and NNC+O2. The NNC+ISO and NNC+O2 groups received bilateral intracerebroventricular injection of NNC 26-9100. The V+O2 and V+ISO groups received equal volume of vehicle control. 24 hours after injection, V+ISO group and NNC+ISO group inhaled 2% isoflurane for 4 hr, while the rest group received 4 hr of 100% O2. Memory was assessed in the Morris water maze. The expression levels of neprilysin(NEP) and insulin degrading enzyme(IDE) in hippocampus were measured by real-time PCR and western blotting. Results Compared with V+ISO group, rats in NNC+ISO group took less time to find platform in day 2 and 3 (P<0.05), spent more time in the target quadrant and crossed more number of platform(P<0.05). There was no significant difference in expression level of NEP and IDE in each group. Conclusion Somatostatin Receptor Subtype-4 Agonist(NNC 26-9100) can improve isoflurane-induced learning and memory dysfunction.

The proliferation of BMSCs is increased following the precondition with low concentration hydrogen peroxide

2016, 36(3):  342-347. 

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Objective To investigate the effect and its mechanism of H2O2 preconditioning-induced the enhancement of anti-apoptosis in BMSCs. Methods BMSCs isolated from rat were treated with H2O2 for 48h, or 50μmol/L H2O2 proconditioning following treatment with 500μmol/L H2O2 combined with or without anti-CXCR7 antibody for 48h. BMSCs viability was measured by MTT assay. The apoptotic cells was measured by Hoechst33342. The expression of SDF-1 and its CXCR4, CXCR7 receptor, Bcl-2, Bax and the key proteins of Akt/mTOR pathway was detected by Western blot. Results 25μmol/L and 50μmol/L was promoted markedly H2O2 BMSCs proliferation and the expression of SDF-1 and its CXCR4, CXCR7 receptor, and key phosphorylated proteins of Akt/mTOR pathway. Transwell migration assay and scraping result were shown that H2O2 preconditioning of BMSCs significantly augmented the migration ability, and protected BMSCs from 500 μmol /L H2O2-induced apoptosis，decreasing the rate of apoptotic cells, but antagonizing by anti-CXCR7 antibody. Conclusion H2O2 preconditioning induces the enhancement of the activity of bone marrow stem cell through CXCR7 receptor.

Association of midtrimester hsCRP with gestational diabetes mellitus

2016, 36(3):  348-352. 

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Objective To explore the associations of high sensitivity C-reactive protein (hsCRP) in the second trimester of pregnancy with gestational diabetes mellitus（GDM）and glucose related traits. Methods 725 women with GDM and 935 women who remained euglycemic throughout pregnancy were enrolled in this study. Pre-pregnancy weight、height and blood pressure were recorded and hsCRP、glucose and insulin levels were detected at 24 to 28 weeks of pregnancy. The associations of hsCRP levels with the blood glucose and insulin levels were analyzed，meanwhile, we also researched the associations of hsCRP with the occurrence of GDM. Results The hsCRP levels in the second trimester of pregnancy were significantly increased in GDM subjects compared with subjects with normal glucose tolerance（NGT）（p<0.01）. Elevated hsCRP was an independent risk factor for GDM（p<0.05）.Fasting blood glucose was associated with hsCRP（P<0.05）;1 h blood glucose after oral 50 grams of sugar was related to hsCRP（P<0.05）; 1，2 and 3h blood glucose after 100 grams OGTT were related to hsCRP（P<0.05）; glycosylated hemoglobin(HbA1c) was related to hsCRP（P<0.05）; fasting plasma insulin was related to hsCRP（P<0.05）;1,2 and 3h true insulin after 100 grams OGTT were related to hsCRP（P<0.05）;HOMA-IR was associated with hsCRP（P<0.05）.Conclusions The elevated hsCRP levels in the second trimester of pregnancy could significantly increase the risk of GDM，which is expected to become one of markers for the diagnosis of GDM.

Effects of Artemis on DNA damage of human hepatic carcinoma cell line BEL-7402/5FU

2016, 36(3):  353-357. 

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Objective To explore the cellular effects of shArtemis on DNA damage of human hepatic carcinoma cell line BEL-7402/5FU in vitro.Methods The experiment were divided into control group，Lipofectamine2000 group，contol plasmid group, shArtemis group;Human hepatic carcinoma cell line BEL-7402/5FU were treated with 0.625,1.25,2.5.5.0 and 10.0μg/mL mitomycin C for 24 and 48 huors.BEL-7402/5FU cell activity examined by MTT and observed the expression of phosphorylated histone2AX (γ-H2AX ) by Western blot.After 48h transfected shArtemis interference plasmid , Western blot detected the quantity of Artemis and γ-H2AX ;Commet assy detect the exent of DNA damage.Result The Determined by MTT show that after 48 h respectively treated within 0.625, 1.25, 2.5, 5.0 and 10.0 μg/mL Mitomycin C, cell activity of liver cancer happen to have significantly decreased.In addition, after 48h by incubating with Mitomycin C,the expression of γ-H2AX increased in a dose-dependent manner;The the expression of γ-H2AX increased following inhibitated Artemis. Moreover the tailing DNA of shArtemis experimental group increased than in the control group.Conclusion mitomycin C can induced DNA damage in human hepatic carcinoma cell line BEL-7402/5FU; shArtemis prmotes DNA damage which induced by mitomycin C.

Relieving effect of substance P on hyperoxia-induced lung injury in premature rats via MAPKs signaling pathway

2016, 36(3):  358-363. 

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Objective: To explore the impact of exposure to hyperoxia on lung tissue from premature rats. Meanwhile, rats were treated with substance P to observe whether it has an influence on hyperoxia-induced lung injury and MAPKs signaling pathway. Methods: Sixty premature Wistar rats were divided randomly into 4 groups: normoxic group， normoxic+SP group，hyperoxic group， hyperoxic+SP group. Each group was divided into 3 subgroups according to the 3 time points of 3d, 7d and 14d. Lung pathology was examined with light microscopy. Wet/dry (W/D) ratio of lung tissue, the content of SP in lung were evaluated. MDA, SOD and GSH-Px were detected. Cell apoptosis was detected by TUNEL. MAPKs was detected by Western blot. Results: The rats in hyperoxia groups had lung injury, and it was increased dependently with the time of exposure to hyperoxia, while treated with substance P the injury could be improved.The W/D ratio of rats in hyperoxia group was significantly increased(P＜0.05), while treated with substance P W/D ratio become reducing(P＜0.05). The content of SP in hyperoxia group was reduced and the content of SP was decreased dependently with the time of exposure to hyperoxia(P＜0.05). The vitality of MDA in the hyperoxia group was increased significantly, and the vitality of hyperoxia+SP group was decreased as compared with that in the hyperoxia(P＜0.05). The vitality of SOD,GSH-Px in the hyperoxia group was decreased(P＜0.05), which become much lower dependently with the time of exposure to hyperoxia, the vitality of SOD,GSH-Px were increased after treated with substance P(P＜0.05). TUNEL-positive cell of hyperoxia groups significantly increased inversely. TUNEL-positive cell was decreased as rats were treated with substance P. The results of Western blotting revealed that the expression of p-JNK,P38 and ERK protein was higher in the hyperoxia group than that in the normoxic group(P＜0.05)，and the expression of p-JNK,P38 and ERK protein increased in time dependently. However, the expression of p-JNK and P38 protein was lessened in hyperoxia+SP group(P＜0.05). The expression of ERK protein was increased in hyperoxia+SP group(P＜0.05). Conclusion: Hyperoxia can lead to lung injury in premature rats and treatment with SP could protect lung injury in oxidative stress condition by inhibiting MAPKs pathway.

miR-30a over-expression inhibits osteogenic differentiation induced by BMP9 in C2C12 cells

2016, 36(3):  364-369. 

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Objective To study the role of miR-30a in bone morphogenetic protein 9 (BMP9) induced osteogenic differentiation of mesenchymal stem cell line C2C12. Methods C2C12 cells were treated with BMP9 conditioned medium to induce osteogenic differentiation. Real-time PCR was used to detect the expression of miR-30 family members in BMP9-induced osteogenic differentiation of C2C12 cells. Effects of miR-30a on BMP9-induced osteogenic differentiation of C2C12 cells were detected by alkaline phosphatase(ALP)，osteocalcin(OCN) and calcium deposition. MTT and cell cycle analysis were used to reveale cell proliferation. Results The expression of only one miR-30 family member, miR-30a, first decreased and then increased during BMP9-induced osteogenic differentiation of C2C12 cells.Over-expression of miR-30a ，which targets on Runx2，led to expression of an early osteogenic marker and a reduction in ALP expression. In addition, we observed decreases in the expression of later osteogenic markers Osteocalcin，as well as calcium deposition. Conclusions BMP9-induced osteogenic differentiation is partially inhibited by miR-30a in mesenchymal stem cell line C2C12.

Targeted blocking LRP increases chemotherapeutic sensitivity to taxol in lung carcinoma cells

2016, 36(3):  370-374. 

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Objective：Using siRNA（Small interference RNA） technology to silence LRP(lung resistance protein),and to explore whether this may improve the sensitivity to taxol in Lung Adenocarcinoma. Method: For building the cell line which is taxol -resistance, the graduated concentrations of taxol was added to the cells. Then the siRNA technology to silence LRP gene in A549/TXL20 was performed.MTT detected the IC50 of taxol after A549/TXL20 transfection. Using qPCR to test the level of expression of LRP mRNA .Western blot detected the protein of LRP in the cells. To build the nude mice model of transplanted tumor by injecting A549/TXL cell into the armpit of nude mice, then detected the function of siRNA aimed at LRP to the resistance of drug in human lung adenocarcinoma cell line, A549/TXL20. Results :After siRNA silencing LRP gene in A549/TXL20 cell line , compared the blank group and the negative group, the control group showed the inhibiting effect of LRP mRNA and protein expression (P<0.01).MTT showed that the sensitivity for taxol in A549/TXL20 was raised.A549/TXL20 cell line after interference could not grow to form transplanted tumors（P<0.01）. Conculsion: siRNA could silence the LRP in A549/TXL20 which is the gene related to Lung Adenocarcinoma resistant drugs, and raise the sensitivity of the cell line to taxol.

Effects of protein kinase B inhibitor Perifosine on ovarian clear cell carcinoma ES2 cell line in vitro

2016, 36(3):  375-379. 

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Objective To investigate the effects of protein kinase B inhibitor Perifosine on ovarian clear cell carcinoma ES2 cell line in vitro. Methods The human ovarian clear cell carcinoma ES2 cells were cultured in vitro and treated with different doses of Perifosine(0、7、9、11、13、15 μmol/L) for 24 h. Survival rate of ES2 cells was evaluated with cell counting kit-8(CCK-8). Morphology of nuclei were observed with DAPI staining. Cell apoptosis was analyzed with Annexin V-FITC/PI apoptosis kit for flow cytometry. The expression levels of Ki-67 and MCM2 mRNA were detected by RT-PCR. Results The survival rate decreased in ES2 cells treated by Perifosine after 24 h(P＜0.05) and the rate was related with concentration to a certain extent. A large number of nuclear fragmentation and apoptotic bodies were observed under fluorescence microscope after treated by Perifosine. Cell apoptosis rate increased with the increase of the concentration of Perifosine(P＜0.01). The expression levels of Ki-67 and MCM2 mRNA were significantly decreased after 48 h(P＜0.01). Conclusions Perifosine has distinctive inhibitory effects on human ovarian clear cell carcinoma ES2 cells, and is able to negatively regulate the expression of Ki-67 and MCM2 mRNA.

Down—regulation of Adipo/AMPK signaling is involved in the pathogenesis of obesity-related asthma

2016, 36(3):  380-385. 

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Objective To investigate the potential effect of adiponectin/ AMP-activated protein (AMPK ) signaling pathway on the airway inflammation and airway hyperreactivity in obesity-related asthma. Methods Forty male C57BL/6 mice were randomly divided into four groups, which were designed as control group(A), asthma group(B) ,obese group(C) and obese asthma group(D). The mice in group A were fed with normal diet , sensitized and challenged with saline. The mice in group B were sensitized and challenged with ovalbumin(OVA) and fed with normal diet. The mice in group C and D were fed with high fat diet, the mice in group D were sensitized and challenged with OVA and the mice in group C used saline instead. After 12 weeks, airway resistances(RL) were determined and the histopathologies of lung tissues were observed .Real-time PCR was used to detect the expressions of adiponectin mRNA、AdipoR1 mRNA and AMPKα mRNA and Westem blotting was used to detect the expressions of adiponectin、phosphor-AMPKα(pAMPKα) and AMPKα proteins．Results The mean weight of high fat food feeding mice was 20% higher than mice feeding with normal diet and achieved the standard of obesity at the 12 weeks after feeding, the difference was significant(P＜0.01). Compared to group A, the expression level of adiponectin mRNA,AdipoR1 mRNA as well as the adiponectin protein and pAMPKα protein in lung were significantly decreased in the other three groups with group D most obviously(P<0.05).Conclusions The airway responsiveness was higher in obesity-related asthma, which may partly due to the down-regulation of Adipo/AMPK signaling pathway.

The characteristics and effects of growth hormone treatment of 16 multiple pituitary hormone deficiency patients with growth hormone deficiency

2016, 36(3):  386-390. 

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Objective: To study the clinical feature and therapeutic effects of 16 patients diagnosed with multiple pituitary hormone deficiency with growth hormone deficiency (GHD). Methods: Clinical data of 16 cases were studied retrospectively. Results: 16 MPHD with GHD patients were included, containing 9 hypothyroidism, 13 hypogonadism and 6 ACTH deficiency. 10 of them had breech or foot presentation. The mean bone age was 11.0±3.5yr, delayed from chronological age, peak GH of L-Dopa provocation test was 0.14±0.17ng/mL. GH dose was 0.11±0.02IU/kg. After GH replacement therapy，the serum concentration of IGF-1 and growth velocity increased. Conclusion: GH replacement therapy is crucial once after one is diagnosed of MPHD excluding the hypothalamus and pituitary lesions. Other pituitary hormones must be replaced sufficiently for MPHD patients. There are no severe adverse events.

Increased expression of miR-221/miR-222 in human thyroid papillary carcinoma

2016, 36(3):  391-392. 

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Expression and significance of ADAM17 in papillary carcinoma of thyroid

2016, 36(3):  393-395. 

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Biological characteristics of HACE1 and advances in malignancy study

2016, 36(3):  396-400. 

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HACE1 is not only an important tumor suppressor gene, which plays an important role in tumor inhibition by mediating cell autophagy, Rac1 ubiquitination and other mechanisms, but also involved in a variety of biological functions about heart protection, anti-oxidative stress and cellular dynamics. Downregulation or mutations of HACE1 gene plays an important role in many human malignancies occurrence, invasion and metastasis process and closely related to prognosis. Therefore, HACE1 may become a new target for cancer therapy, providing new directions and opportunities for treatment of malignant tumors.

Progress in molecular mechanism of resveratrol-mediated reversal of tumor drug resistance

2016, 36(3):  401-405. 

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Resveratrol is a kind of natural plant extract with low toxicity. In recent years, studies have shown that resveratrol can enhance the sensitivity of resistant cells to chemotherapy drugs and significantly reverse the Multi-drug resistance(MDR) of tumor by mechanisms such as ATP-binding cassette transporter family, signaling pathways, SIRT1, induction of apoptosis and so on , which will provide a new thought to solve the problem of MDR. Above all, resveratrol is respected to become a novel MDR reversal agent.

The clinical application of EEG in Parkinson's disease research

2016, 36(3):  406-410. 

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Parkinson's disease (PD) is a common neurodegenerative disease in middle-aged and elderly population. PD patients with observable symptoms suffer from irreversible degeneration of neural system, but the treatment and prognosis are challenging. Electroencephalography (EEG) is an important approach in neuroscience research, which could record electrical signals from the entire brain with high temporal resolution, and it is widely applied to clinical PD research. This review focuses on EEG investigations on non-motor symptoms, motor symptoms, and treatment efficacy of PD. The advantages and limitations of EEG technique in clinical applications are also discussed.

Progress in research of the role of lncRNA in gliomas

2016, 36(3):  411-414. 

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The latest study that long non-coding RNAs(lncRNA), regarded as a new class of non-coding gene regulators, can regulate gene expression in 3 aspects. The changes of the hereditary material can affect lncRNA gene expression and lncRNA may contribute to the proliferation and apoptosis, the invasiveness and differentiation of glioma cells, and to angiogenesis. LncRNA can serve as novel biomarkers and therapeutic targets to help diagnose and treat gliomas.

Development of the Willed Body Donation Program promotes anatomical teaching reform

2016, 36(3):  415-418. 

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Based on a strong willed body donation registration and receptionprogram, the number of donation for medical education purposes has been increasing steadily during the past decade. Willed body donation not only provides a legal, consistent and high-quality source of cadaver for anatomical teaching, but also expands the scope of anatomy into clinical training and research. The spirit of donation has rejuvenated the traditional anatomical teaching via introducing humanity, creative thinking and clinical collaboration, all of which have significantly motivated the medical students.

The application of 3D visualization touchscreen table in standardization of surgical training: An analysis of faculty training

2016, 36(3):  419-423. 

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Objective To explore 3D multi-touch visualization touchscreen table (MVT), this novel tool in standardization of surgical training, and to analyze its features and prospects in surgical teaching. Methods During faculty training seminars for standardized residency program, the educators were trained with how to learn surgical cases via this novel tool MVT. Then, they completed the survey, and through the scoring system, with 5 being the highest rating and 1 being the lowest evaluation, data was collected for further statistical analysis. Results A total of 106 educators participated in this session, aging 24-65 years old. Before our demonstration, neutral attitude were held by educators in apply MVT into surgical teaching (3.41 ± 1.44), while after the training given, educators obviously considered using this table into clinical teaching (4.25 ± 0.84). Significant difference was found between them (p <0.001). After the demonstration, educators felt that this table can be used simpler and easier to clarity a complex anatomical structure. Besides, real cases on the table could be very helpful for learning. Moreover, most educators held its useful information for them to improve surgical residency, with scores above 4.20. Conclusion Continuous application of 3D visualization of multi-touch technology could facilitate surgical trainees with good imaginal thinking and diagnostic thinking. This novel tool would further affect the future mode of teaching and diagnosis, and be helpful to residency training.

The application of MOOC in medical humanities education

2016, 36(3):  424-426. 

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MOOC(Massive Open Online Course)as a new type of online education model has been widely used in teaching. This article through present situation of medical humanities to explore how to apply MOOC in medical humanities education and analysis to put forward challenges in the development of medical humanities .The author believes that through MOOC of the modern medical humane education reform, so that the medical curriculum rationalization and medical teachers and students get more high-quality resources in order to promote the development of medical humanities, is worthy of further discussion topic.

Use of TTE simulation in teaching basic Transthoracic Echocardiography skills to anesthesiology residents

2016, 36(3):  427-432. 

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Objective To assess the impact of simulation-based Transthoracic Echocardiography (TTE) training versus traditional lecture-based training on basic TTE skills in anesthesiology residents; Methods In this prospective randomized study, 42 anesthesiology residents (in anesthesia clinical training years 1 to 3)were randomized to either control (n = 21) or simulation groups (n = 21) for TTE training. A standardized pretest was administered before TTE training sessions of 40 minutes each. The training session used a lecture-based video didactic in the control group or a TTE simulator in the simulation group. Comprehension in both groups was then assessed using a written post test and by performing a TTE examination on a volunteer subject. TTE examinations were graded on the ability to acquire the correct image, image quality, anatomy identification, and time required to attain proper imaging ; Results Pretest scores revealed similar knowledge among residents . The simulation group scored higher on all criteria after the training session: written post test (43.3%±10.8% VS 52.6%±17.6%，P<0.05), volunteer subject post test image quality scores (0 to 25 scale) (7.7±5.1 VS 17.1±4.8，P<0.001), anatomy identification scores (0 to 25 scale) (11.0±7.3 VS 18.2±6.3；P=0.001); Conclusion This prospective randomized study demonstrated that anesthesiology residents trained with simulation acquired better skills in TTE image acquisition and anatomy identification on volunteer subjects.