Table of Content

05 May 2016, Volume 36 Issue 5

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The oxidative stress induced by glucose oxidase suppresses the current of α4β2 nicotinic acetylcholine receptor

2016, 36(5):  577-580. 

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Objective To investigate the effect of oxidative stress induced by glucose oxidase (GO) on the current of α4β2 nicotinic acetylcholine receptor (nAChR). Methods HEK293T cells were co-transfected with plasmid α4 and β2 after 48 h. Experimental groups were divided into control group, GO group and CAT group. To measure the level of reactive oxygen species (ROS), we used confocal microscopy to detect fluorescence intensity by DCFH-DA fluorescent probe; the currents of α4β2 nAChR were detected by whole-cell patch clamp. 1 μmol/L, 10 μmol/L and 30 μmol/L ACh were used to detect the receptor function. 10 μmol/L ACh was used to detect current changes. Results The currents from α4β2 nAChR in HEK293T cells were increased in a dose-dependent manner; the treatment of 3.5 kU/L GO for 1 h significantly incresed the level of ROS in HEK293T cells (P <0.001); with the consecutive application of ACh, the currents in control group were still stable after 10 times administration, while the GO-induced oxidative stress could gradually lead to the currents of α4β2 nAChR run-down (P <0.01); the 450 kU/L of catalase (CAT) could prevent the currents running down (P <0.01). Conclusion The GO-induced oxidative stress could cause α4β2 nAChR currents gradually run down. ROS may play an important role in the current run-down of α4β2 nAChR.

Effect of E3 ubiquitin ligase RNF121 on expression of proinflammatory cytokines in LPS-treated macrophages

2016, 36(5):  581-585. 

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Objective To estimate the effect of E3 ubiquitin ligase RNF121 on the expression of proinflammatory cytokines in LPS-stimulated macrophages. Methods The level of RNF121 in mouse peritoneal macrophages treatment with LPS was detected by Weston-Blot. RNF121 expression was reduced by RNA interference (si-RNF121). After LPS stimulation, the mRNA levels of TNF-α and IL-6 in mouse peritoneal macrophages were detected by real-time quantitive PCR (q-PCR). The levels of TNF-α and IL-6 in supernatants were detected by ELISA. The level of phosphorylated p65 (p-p65) was detected by Weston-Blot. The NF-κB activity was measured by dual-luciferase assay. Results After treatment with LPS, the RNF121 protein level was significantly decreased (P<0.05) in macrophages. However, protease inhibitor MG132 can significantly increase the RNF121 protein level. Decreased the expression of RNF121 by si-RNF121 and stimulated by LPS in macrophages, TNF-α and IL-6 expression levels (mRNA and protein levels) were significantly decreased (P<0.05), and the transcription factor phosphorylated p65 (p-p65) was also significantly decreased (P<0.05). In addition, NF-κB activity was also decreased (P<0.05). Conclusions The E3 ubiquitin ligase RNF121 was involved in the regulation of the expression of TNF-α and IL-6 in innate immune responses in mouse macrophages by NF-κB pathway.

Livin promotes epithelial–mesenchymal transition of SGC7901 cells through activation of AKT signaling

2016, 36(5):  586-589. 

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Objective To investigate whether livin expressed in gastric cancer SGC7901 cells could affect epithelial–mesenchymal transition (EMT). Methods SGC7901 cells were infected with livin shRNA lentivirus, stable transfection cells were selected. The invasion and migration ability of the stable transfection cells were detected by Transwell and Wound healing assay. The protein of E-cadherin and Vimentin were detected by Western blot. The activation of Akt signaling was detected by Western blot. Results Silencing livin in SGC7901 cells could inhibit the invasion and migration ability in vitro(P＜0.05). The protein of E-cadherin was significantly increased and vimentin was significantly decreased when livin was Silenced by livin shRNA(P＜0.05). Also, the activation of Akt signaling was significantly decreased(P＜0.05). Conclusion Livin expressed in gastric cancer cell promotes invasion and migration ability in vitro through induction of EMT by activation of AKT signaling.

Xinkeshu and fluvastatin inhibit adhesive molecules expression and inflammation in plaque of atherosclerosis in rabbit

2016, 36(5):  590-593. 

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Objective: To observe the effect of Xinkeshu (XKS) and fluvastatin on adhesive molecules and inflammationin in plaque of atherosclerosis in rabbit.Methods: The New Zealand white rabbits were randomly divided into four groups: control group, high-fat group, fluvastatin group and Xinkeshu(XKS) group (9 /per group).The atherosclerotic plaques were evaluated by HE staining. Expression of MMP-9,macrophage andα-SMA in atherosclerotic plaques were detected by immunohistochemistry. The SOD and MDA were also measured. Results：Compared with high-fat group，the expressions of VCAM-1 in atherosclerotic plaques significantly decreased with the treatment of XKS and fluvastatin(P<0.01). The expression of MMP-9，macrophage infiltration andα-SMA were also lower in the XKS group and fluvastatin group than that in high-fat group (P<0.01)，while serum SOD activity increased (P<0.01). Conclusions： XKS and fluvastatin play an important role in the protection of endothelial function and anti-atherosclerosis effect by its anti-inflammatory and anti-oxygen radical.

Genetic asymmetry in Type II CD36 deficiency

2016, 36(5):  594-597. 

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Objective To use DNA sequencing methods to determine the type II CD36 deficiency genotype, and to analyze the genetic of CD36 gene in guangzhou blood donors. Methods Using PCR-SBT technology to analyze 998 blood donors in guangzhou CD36 genetic coding region; To analyze CD36 protein expression level by flow cytometry instrument. Results In 998 subjects, subject to wild type gene, a single mutation and two mutations was 980, 13 and 5 respectively. Type I CD36 deficiency The type I CD36 deficiency is most commonly detected in subjects who are homozygous or compound heterozygous for mutations. Conclusion Monocyte CD36 protein may be influenced by genetic dose dependent; On the other hand, can be affected by genetic factors besides the encoding gene, resulting in genetic asymmetry in type II CD36 deficiency.

miR-92b-3p promotes the osteogenesis of humans umbilical cord mesenchymal stem cells

2016, 36(5):  598-603. 

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Objective To investigate the functional role of the miR-92b-3p in osteogenesis of human umbilical cord mesenchymal stem cells (hUCMSCs). Methods The expression level of miR-92b-3p after transfection of synthetic miR-92b-3p mimic was confirmed by qRT-PCR. The transfected hUCMSCs were induced to osteoblast differentiation. Alizarin red S staining was used to determine osteogenic ability of different transfected cells. qRT-PCR was used to analyze the osteogenic marker gene expressions. Using bioinformatics analysis to predict the potential target of miR-92b-3p, and the dual luciferase report gene system and Western blot to verify the predication. Results The expression of miR-92b-3p in the process of osteodifferentiation was significantly higher than that of the control group (P < 0.01). Overexpressed miR-92b-3p could promote hUCMSCs osteogenetic ability. While inhibiting miR-92b-3p could reduce the osteogenetic differentiation level of hUCMSCs. Overexpressed miR-92b-3p could significantly reduce the expression of DKK1, while inhibition of miR-92b-3p could significantly improve the expression of DKK1. Conclusion: MiR-92b-3p can promote the osteogenesis of hUCMSCs by downregulating the DKK1.

RAGE inhibitor FPS-ZM1 reduced isoflurane anesthesia induced cognitive dysfunction in aged rats

2016, 36(5):  604-609. 

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Objective To investigate the effect of FPS-ZM1 on isoflurane anesthesia induced cognitive dysfunction in aged rats. Methods Sixty aged rats were randomly divided into four groups(n=15)as follow: NS+O2、NS+ISO、FPS-ZM1+ISO、FPS-ZM1+O2.The NS+O2 and NS+ISO groups received intracerebroventricular injection of normalsaline 10 μL. The FPS-ZM1+O2 and FPS-ZM1+ISO groups received intracerebroventricular injection of FPS-ZM1 10 μL(5 g/L). 24 hours after intracerebroventricular injection, The NS+O2 and FPS-ZM1+O2 groups inhaled a gas mixture of oxygen and air for 4 h, and the NS+ISO and FPS-ZM1+ISO groups inhaled 2% isoflurane for 4 h. The tests of learning and memory were performed in Morris water maze, RAGE and amyloid β protein(Aβ) expression in hippocampus CA1 area was determined by immunohistochemistry. Results Compared with NS+ISO group, the escaping latency significantly shortened from 1 to 4 day, and the probe time was significantly increased, the time of staying at the original platform quadrant was significantly longer inthe FPS-ZM1+ISO group. In addition, the level of RAGE and Aβ expression of groups FPS-ZM1+ISO in hippocampus CA1 area was significantly reduced. Conclusion FPS-ZM1 may improve the process of spatial learning and memory by inhibiting Aβ expression in hippocampus CA1 area of aged rats.

miR-551b-3p decreases in gastric cancer tissue and inhibits the proliferation, migration and invasion of gastric cancer cell line HGC-27

Di ZHAI Fang WANG Jia YU

2016, 36(5):  610-614. 

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Objective To investigate the expression and the expression change of miR-551b-3p in gastric cancer and its effect on the function of gastric cancer cells. Methods miR-551b-3p was detected by RT-PCR in 60 cases of gastric cancer samples and their paired adjacent normal control tissues. HGC-27 cell line was infected by miR-551b-3p mimic, then CCK-8 assay was used to determine the influence of miR-551b-3p on HGC-27 cell proliferation; Wound-healing assay was used to determine cell migration; Transwell assay was used to determine cell invasion. Results 1) The expression of miR-551b-3p in gastric tumor tissues was significantly lower than that of the adjacent tissues (P<0.05). 2) HGC-27 cell proliferation, migration and invasion were significantly decreased while the cell infected by mimic-miR-551b-3p. Conclusions miR-551b-3p expresses lower in gastric cancer tissues and is able to inhibit gastric cancer line HGC-27 proliferation, migration and invasion. miR-551b-3p is closely associated with the development of gastric cancer.

Human mesenchymal stem cells promote breast cancer cells migration and invasion by co-culture in vitro

2016, 36(5):  615-620. 

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Objective To study the effect of human bone marrow derived MSCs (hBM-MSCs) on the migration and invasion capacity of breast cancer cells in co-culture system. Methods Cytokines secreted by BM-MSCs were detected using ELISA assay. Tumor microenvironments were mimicked by co-culturing of BM-MSCs with breast cancer cell lines (MCF-7 and MDB-MA-231) separately. Capacity of migration and invasion were compared after co-culture using Transwell system．Activation of signal transducer and activator of transcription 3 (STAT3) was detected by Western blot. Experession of metastasis-associated gene matrix metalloproteinases (MMP2 and MMP9) were detected by realtime RT-PCR. Result HGF, IL6, VEGF and TGF β 1 were detected in the medium of BM-MSCs. Co-culturing with BM-MSCs can significantly enhance the migration and invasion capacity of breast cancer cells (the migration and invasion of MCF-7: P<0.01; the migration of MDA-231: P<0. 05; the invasion of MDA-231: P<0. 05)．Signaling pathway of p-STAT3 and p-ERK were activated dramatically(P<0.01), and the expression of MMP2 and MMP9 were increased markedly(P<0.01). Conclusions BM-MSCs may promote the migration and invasion capacity of breast cancer cells by indirect co-culture of the two cell lines．

The effects of exosomes from adipose-tissue mesenchymal stem cells on the expression of microRNAs in MCF7 cells

2016, 36(5):  621-626. 

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Objective To evaluate the biological characteristics of exosomes secreted by adipose-tissue derived mesenchymal stem cells(AD-MSCs) and to evaluate the effects of these exosomes on microRNA expression of breast cancer cell MCF7. Methods Exosomes were isolated from supernatant of AD-MSCs by ultracentrifugation and ultrafiltration. The morphology was observed under electron microscope. Surface marker expression of exosomes was detected by Western blot. Dil-labeled exosomes were added into MCF7 culture medium and the internalization of exosomes by MCF7 was observed under fluorescence microscope. MicroRNA array chip was used to investigate the microRNA expression changes in MCF7 before and after exosome treatment for 24 h. MicroRNA array chip was also used to detect the expression of microRNAs in exosomes. Results AD-MSCs secreted 30~100 nm exosomes which expressed protein makers CD63 and HSP70. Exosomes from AD-MSCs could be internalized by MCF7 and cause microRNA expression changes in MCF7. 244 microRNAs were differentially expressed in MCF7 after treatment with exosomes for 24 h(174 were up-regulated and 70 were down-regulated, with fold change>2). MicroRNA array analysis of exosomes indicated that most of the increased microRNAs in MCF7 were generated endogenously, not derived from exosomes. Conclusion Exosomes from AD-MSCs can regulate microRNA expression of breast cancer cell MCF7, which might be the new mechanism by which AD-MSCs affect MCF7.

Dendritic projections of motor neurons in the rat spinal cord anterior horn associated to the large intestine channel of hand-yangming and the pericardium channel of hand-jueyin

2016, 36(5):  627-632. 

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Abstract: Objective To map the specific distribution of the dendritic projections of the motor neurons related to the large intestine channel of hand-yangming and the pericardium channel of hand-jueyin in the spinal cord of rat. Method Cholera toxin B subunit combined with the horseradish peroxidase (CB-HRP) was injected as a neuronal tracer into the acupoints in the muscles and deep tissues of large intestine channel of hand-yangming and the pericardium channel of hand-jueyin. Each acupoint received an injection of 5 μL CB-HRP. Three to four days after injection, rats were perfused with fixatives and the cervical spinal cord was harvested, followed by TMB chemo-histological reactions to label the motor neurons. CB-HRP positive motor neurons?were observed with light microscope in the transverse and the horizontal sections.?Result?The motor neurons innervating the muscle groups related to each of the two given meridians (the large intestine channel of hand-yangming and the pericardium channel of hand-jueyin) formed a longitudinal?neural column?in the ventral horn of the cervical spinal cord.?Dense dendritic-dendritic and dendritic-soma projections were observed between the motor neurons within the same neural column. Conclusion?Our results indicated that the motor neurons related to a given meridian formed a longitudinal motor neuron column in the spinal cord ventral horn. The dendritic-dendritic and dendritic-somal projections?exclusively within the same motor neuron column corresponding to a certain meridian channel might be the morphological basis of meridian propagation as well as meridian reflex activities.?

Effect of decompressed on demyelination and BDNF expression in spinal cord of rats after compressed spinal cord injury

2016, 36(5):  633-637. 

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Objective To observe the influence of brain-derived neurotrophic factor on demyelination of myelinated nerve fibers after compressed spinal cord injury, so as to build up the experimental basis for BDNF’s repairing function on nerve fibers demyelination. Methods rats were divided into groups averagely： normal group, sham group, compressed group and decompressed group. The CSCI mode was established by a custom-made compressor. For the compress group, the rats were subjected to spinal cord compression for 12 h; for the decompress group, only 1h; while the sham group just exposed the spinal cord .The changes of nerve fibers were observed on day 1,3and7 after CSCI by osmic acid staining. The expression of BDNF and MBP were detected by Western blot and double-labelling immunofluorescence.Result axonal demyelination occurred in both compress and decompress group. Myelin sheath became swelling, degenerating and breaking down with the flowing of time. The expression of BDNF and MBP decreased gradually as the time went by(p<0.05). But compared to the compress group, the demyelinating in the decompress group was less serious and the reduction both in MBP and BDNF was less and slower(p<0.05).Conclusion early decompression can reduce demyelination which may be related with the expression of BDNF in CSCI.

Expression of miR-363-3p in gastric cancer tissue and its effect of proliferation, migration and invasion on gastric cancer cell line HGC - 27

2016, 36(5):  638-643. 

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Objective To investigate of the expression changes of miR-363-3p in gastric cancer and its functional significance in HGC-27 cell line. Methods Real-time PCR analysis was used to detect the relative expression of miR-363-3p in 59 pairs of gastric cancer tissues and their normal control tissues. miR-363-3p mimics was transfected into HGC-27 cell line for miR-363-3p overexpression. The effects of miR-363-3p overexpression on cell proliferation, migration and invasion were detected by CCK-8, wounding heal and Transwell assays. Results Overexpression of miR-363-3p in gastric cancer cell line （HGC-27）. Proliferation text results showed that miR-363-3p could inhibit cell proliferation. Scratch text results showed that miR-363-3p could inhibit cell migration. Transwell text results showed that miR-363-3p could inhibit cell invasion. Conclusions miR-363-3p functions as a tumor suppressor miRNA in gastric cancer. Therefore it might have the potential to be a new target for gastric cancer therapy.

Over expression of c-Met protien mediats EPCs distribution in lung of hypoxia pulmonary arterial hypertension rat

2016, 36(5):  644-650. 

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Objective To discuss overexpression c-Met protein mediated distribution of EPCs in HAPH model.Methods To establish the model of HPAH,primary culture rat bone marrow-derived EPCs and identificated, adenoviral with c-Met gene infected EPCs and Cells were divided into control group,viral load group,c-Met over-expression group.c-Met expression in protein and gene level were detected by Western Blot and Real time PCR , endothelial progenitor cells were marked by CM-Dil , the cells were infused through the tail vein of rat,distribution of the cells were observed in of the major organs afte 1W. Results c-Met mRNA expression of control group、viral load group and c-Met over-expression group were 0.19，0.17 and 1.85；c-Met protein expression were 0.24、0.17 and 0.82. The positive rate of EPCs in lungs、liver、spleen and kideny of EPCs group were 1.7 ％,0.5 ％,7.9 ％ and 9.4 ％；The positive rate of EPCs in viral load group were：1.5 ％,0.4 ％,8.3 ％ and 8.5 ％；The positive rate of EPCs in c-Met EPCs group were 9.3 ％,0.2 ％,1.3 ％ and 0.3 ％,The positive rate of EPCs in c-Met EPCs group significant higher in lung and decreased in spleen and kideny compared with control group（P＜0.001）.Conclusions Over expression of c-Met protein make EPCs superiority distributed in lung of HPAH model.

Increases the circulating smooth muscle progenitor cells in acute spinal cord injury mice

2016, 36(5):  651-655. 

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Objective In this report, the absolute SMPCs count was performed by two single-platform technologies. The reliability of two methods, including intra- and inter-observer variability, was assessed using the intra-class correlation coefficient (ICC), respectively. The stability of SMPCs numbers in blood samples was analyzed. Methods The blood samples were obtained from Institute of Cancer Research (ICR) mice. The samples were processed through now lyse/one-wash procedure and c-kit-/CD140b+/lineage-/sca-1+ SMPCs were analyzed by exclusion of dead cells and by fluorescence-minus-one control. SMPCs were enumerated by two methods: 1) bead-based 123count eBeads? (eBioscience) Count; 2) ‘direct volume’-based Accuri C6 Flow Cytometer? (BD) Count. The cells were measured immediately and after storage of blood samples for 24 h and 48 h, respectively. The SCI in mice were induced by Allen’s device and the number of SMPCs was detected 24h after SCI. Results There are good intra-observer and inter-observer agreement in both methods. It was demonstrated SMPCs are unstable in blood samples. The number of SMPCs increased after 24h in spinal cord injury (SCI) mice. Conclusions Two reproducible protocols for SMPCs quantification were established. Acute spinal cord injury increases the smooth muscle progenitor cells in mice. These protocols should facilitate future studies to further define the role of SMPCs as cellular biomarkers in SCI.

Diagnostic value of combined test of serum autoantibodies to tumor related antigens in non-small-cell lung carcinoma

2016, 36(5):  656-660. 

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Objective To investigate the diagnostic value of single and combined test of seven serum autoantibodies to tumor related antigens (NY-ESO-1，p53，CDK2, AnnexinA2, IMP1, Ubiquilin1 and PGAM1) in non-small-cell lung carcinoma. Methods Serum levels of seven autoantibodies in NSCLC patients (48) and health controls (45) were measured simultaneously by multiplexed Luminex assays. Receiving operating characteristic (ROC) curves were used to analyze the diagnostic accuracy in single and combined tests. Results 1) The level of seven autoantibodies in serum were higher in patients with NSCLC than that in health controls (p<0.05)； 2) The analysis of ROC curves showed that the AUC of NY-ESO-1 and p53 were more than 0.7, while the sensitivities of AnnexinA2 was 88.89 %； 3) The AUC, sensitivity and specificity of combined assay of seven autoantibodies for NSCLC were 0.812, 68.75 % and 91.11 %. Conclusions All these seven autoantibodies have certain significance in the diagnosis of NSCLC and combination of seven autoantibodies can enhance the diagnostic accuracy for NSCLC.

Cyclopamine relieves neuropathic pain of rats by Inhibiting SHH signaling pathway

2016, 36(5):  661-665. 

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Objective To explore the effect of inhibiting SHH signaling pathway on the change of hyperalgesia and the change of the BDNF, NR2B level in spinal cord of spared nerve injury rats. Methods Rats were randomly divided into Sham operation group (control group), Spared nerve injury group (SNI group), Solvent group group(SNI-vehicle group) and inhibitor group (SNI-CP group) (n=12). Adopt the method of sciatic nerve branch selective damage to make neuropathic pain models. Pain behavior was assessed and mechanical pain threshold(PMWT) was measured at 1d before operation and the 1, 7 d after SNI operation. After 7 d, animals were then sacrificed and the lumbar segement (L4-6) of the spinal cord was obtained. The expression of SHH, GLI1 and BDNF was detected by Western blotting and Quantitative Real-time PCR method, and the expression of NR2B in the spinal cord was detected by immunohistochemistry method. Results Compared with control group, after the SNI operation in SNI group, SNI-vehicle group and SNI-CP group PMWT was significantly decreased, the protein and mRNA expression of SHH, GLI1 and BDNF were obviously increased, the protein expression of NR2B were obviously increased(P＜0.05). Compared with SNI group, PMWT was significantly increased in SNI-CP group, the the protein and mRNA expression of SHH, GLI1 and BDNF were partly decreased, the protein expression of NR2B were partly decreased(P＜0.05). Conclusions Inhibition of SHH signaling pathway can alleviate neuropathic pain in SNI rats and its mechanism may be related to the regulation of BDNF, NR2B in the spinal cord level.

Role of mBDNF /Akt/CREB and proBDNF/RhoA signaling imbalances in cognitive dysfunction caused by propofol in neonatal rats

2016, 36(5):  666-671. 

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Objective To investigate the effect of mBDNF /Akt/CREB，proBDNF/ RhoA signaling imbalances in cognitive dysfunction caused by propofol in neonatal rats Methods Rats aged 7 days weighing 12-16g were randomly divided into 6 groups(n=12 each): group C were intraperitoneally injected with 0.9 % normal saline for 7 days, group P1 were injected normal saline for 6 days and propofol on the 7th day, group P2 were recived propofol for 7 days, group T，K and D were given propofol for 7 days and then respectively injected with TAT-Pep5，K252a and DMSO.Blood gas analysis and blood glucose were detected of 6 rats in each group. The other 6 rats in each group were subjected to Morris water maze to access learning and memory functions at age of 25 d and decapitated after the tests.The hippocampal tissues were taken to measure mBDNF /Akt/CREB，proBDNF/ RhoA by Western blot.Results Compared with group C, the escape latency was prolonged（P＜0.05）, space exploration time was shortened in group P1 and more obvious in P2. Compared with group D, the escape latency was prolonged, space exploration time was shortened in group K（P＜0.05）, the escape latency was shortened, space exploration time was prolonged in group T（P＜0.05）. The expression of mBDNF /Akt/ CREB in hippocampus were decreased in group P1 and gradually decreased in group P2（P＜0.05）. The proBDNF/ RhoA in hippocampus were increased in group P1 and gradually increased in group P2（P＜0.05）.Compared with group D, the Akt/ CREB were down-regulated in group K, the RhoA were reduced in group T（P＜0.05）. Conclusions Propofol anesthesia supressed the anti-apoptosis signal pathway of mBDNF /Akt/ CREB and enhanced the apoptosis signal pathway of proBDNF/RhoA in the hippocampal in neonatal rats, which may be the mechanism by which propofol leads to long-term cognitive dysfunction.

Effect of miR-135a expression on the sensitivity of paclitaxel in gastric cancer cells

2016, 36(5):  672-677. 

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Objective To investigate the relationship between miR-135a expression and paclitaxel sensitivity in gastric cancer cells, and explore the possible mechanisms. Methods The differential microRNAs were screened using Taqman microRNA assay in the serum of gastric cancer patients with different efficacies treated with paclitaxel chemotherapy. microRNA-135a (miR-135a), which had relationship with paclitaxel sensitivity, was selected in the following study. The influence and possible mechanisms of miR-135a expression on the sensitivity of paclitaxel in gastric cancer cells were investigated by using cell proliferation curve, flow cytometry, and Western blot. Results The expression level of miR-135a in patients with progressive disease treated with paclitaxel was significantly higher than that in patients with partial response（P<0.05）, suggesting its relationship with paclitaxel resistance. miR-135a could decrease the sensitivity of paclitaxel in gastric cancer cells through inhibiting cell cycle arrest at G2 phase and cell apoptosis induced by paclitaxel（P<0.05）. Conclusions miR-135a might be a predictive marker for paclitaxel efficacy in future clinical practice.

α-Lipoic acid reduces TNF-α release of LPS-induced Raw264.7 cell

2016, 36(5):  678-681. 

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Objective To investigate the effect of α-lipoic acid on the release of TNF-α of LPS-induced Raw264.7 cell. Methods The Raw264.7 cell were pretreated with ALA or ALA and ERK /NF-κB inhibitors or activators for 2 h, and then stimulated with LPS (1 mg/L). The levels of tumor necrosis factor (TNF-α)in cultured supernatant were determined by ELISA kit．T-pERK,pERK and NF-κB were analyzed by Western blotting．Results ALA inhibited LPS-induced release of TNF-α（p<0.05),while the effect was reversible when the Raw264.7 cell were pretreated with ALA and ERK /NF-κB activators（p<0.05).ALA inhibited the phosphorytion of ERK and NF-κB （p<0.05),while the effect was strengthen when the Raw264.7 cell were pretreated with ALA and ERK /NF-κB inhibitors（p<0.05).Conclusions α-lipoic acid attenuates LPS-induced release of TNF-α by ERK and NF-κB signaling pathway．

Construction of stably expressing TRIM22 HEK293T cell line and its role of inhibiting influenza virus replication

2016, 36(5):  682-687. 

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Objective To construct stably expressing TRIM22 HEK293T cell line and investigate the inhibitory function of TRIM22 to influenza A virus replication. Screen the IAV proteins which can interact with TRIM22. Methods Human TRIM22 gene was cloned onto pQC-XIP with FLAG tag using PCR. Then, the recombinant plasmid and pVSV-G, pGag-Pol were co-transfected into HEK293T cells for packaging retrovirus. Packaged retrovirus was collected and used to infect HEK293T cell, followed by screening with Puromycin. Finally, expression of TRIM22 was detected by Western blot. The inhibitory function of TRIM22 to influenza virus replication was detected with IAV-LUC. The interactions between IAV proteins and TRIM22 were detected by BiLC. Results Gene sequencing indicated that pQC-XIP-TRIM22 was successfully constructed. TRIM22 protein was detected in lysates of stable cell line. Contrast to stable cell line expressing EGFP, that expressing TRIM22 could inhibit the Influenza virus replication. TRIM22 can bind to NP. Conclusions Stable cell line expressing TRIM22 was successfully constructed. TRIM22 can inhibit the influenza virus and it can interact with NP.

Identification of human GIPC2 core promoter and potential cis-acting element using sequence homology approach

cheng zhikai

2016, 36(5):  688-693. 

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Objective To identify human GIPC2 core promoter and potential cis-acting element using sequence homology. Methods Homologous sequence of GIPC2 was identified by comparing GIPC2 coding and non-coding sequence from different species in Vista database. A series of luciferase constructs comprising human or rat GIPC2 sequences truncated at various lengths upstream of translation initiation site were tested in HEK293T and PC12 to confirm the core promoter sequence. Conserved intronic GIPC2 sequences were each inserted upstream of human core promoter to test for potential cis-acting regulatory activities. Results The human GIPC2 core promoter sequence spans from +32 to +190 upstream of translation initiation site. A homologous sequence 1500bp upstream of human GIPC2 translation initiation sites represses GIPC2 promoter activity and a homologous sequence from the first intron promotes GIPC2 promoter activity. Conclusions we have identified human GIPC2 core promoter and potential cis-acting element using sequence homology.

Improvement of bone mineral density in patients with Turner syndrome treated with early estrogen replacement therapy

2016, 36(5):  694-697. 

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Objective To explore the effect of early estrogen replacement therapy on bone mineral density (BMD) in patients with Turner syndrome（TS）.Methods 27 TS patients were selected from Nanism Clinic of Endocrinology Department of Peking Union Medical College Hospital from August 2005 to August 2015. Heights and weights were measured in all patients. The BMD was measured by dual-energy X-ray absorptiometry(LUNAR-PRODGIY Type).We compared the BMD before and after estrogen replacement therapy in patients with TS and the effects on BMD of the starting age of estrogen treatment. Results Estrogen treatment after one year, 1～3 years,> 3 years ,L2-4 BMD were (0.96±0.13) g/cm2、(0.92±0.13) g/cm2 and( 0.93±0.14) g/cm2 respectively, significantly higher than baseline(0.73±0.08) g/cm2、(0.70±0.13) g/cm2 and (0.75±0.07) g/cm2.The BMD L2-4 of whose estrogen therapy was initiated < 18 years group (0.96±0.14) g/cm2was significantly higher than the girls who started estrogen therapy ≥ 18 years group (0.90±0.10) g/cm2.Conclusions Early estrogen replacement therapy can effectively improve BMD in patients with TS, but other factors affecting BMD still need to be further explored.

High incidence of abnormal liver function and the affecting factors in patients with Turner syndrome

2016, 36(5):  698-701. 

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Objective To investigate the incidence of abnormal liver function and the affecting factors in Turner syndrome (TS). Methods Forty-four TS patients aged 13.6±5.0 years old were recruited from the Department of Endocrinology, Peking Union Medical College Hospital during March 2010 to July 2015. And 120 girls aged 13.2±0.7 years old were selected as normal controls. Height, weight and liver function tests including alanine transaminase (ALT) and total bilirubin were measured in all 164 participants. All 44 TS patients were confirmed with chromosome karyotypes. Results The incidence of abnormal liver function in TS patients was 18.2%, mainly manifested as ALT elevation, while the incidence of abnormal liver function in control group was 3.3%. There were no significant differences in age, BMI and karyotypes among TS patients with or without ALT abnormality. Conclusions The incidence of abnormal liver function is higher in TS patients. Regular liver function tests should be performed in these patients. And the diagnosis of TS should be considered when abnormal liver function was noted in female patients with short stature.

The new understanding in functions of angiogenesis in tumor progression and organ fibrosis

2016, 36(5):  702-705. 

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The microvascular density is significantly associated with tumour metastasis and organ fibrosis, angiogenesis can be the common target for the therapy of tumor and organ fibrosis. However, traditional anti-angiogenesis strategies attempt to reduce the tumour vascular supply, but their success is restricted by insufficient efficacy or development of resistance. Moreover, the potential angiogenic therapies with the primary aim of ameliorating interstitial fibrosis is falling into bottleneck. Because it is strongly associated with inflammation and changes in vascular permeability. In this review, we will provide a brief review of current understanding of endothelial cell metabolism and vascular niche to understand the underlying relationship between tumor progression and organ fibrosis in angiogenesis.

Research progress of CAR-T cells in tumor clinical trials

2016, 36(5):  706-710. 

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Currently, chimeric antigen receptor, CAR-related cellular immunotherapy is venerated as the gorgeous star in tumor biotherapy. The CAR-T cell such as which targeted CD19 has obtained the good curative effect in conquering B lymphocyte malignancies. The latest CAT-T related references were collected and summarized to mainly analyse the data of clinical trial, clinical effects and potential adverse reaction treated with CAR-T.

Investigation of the tutorial system for residents training in diagnostic ultrasound speciality

2016, 36(5):  711-714. 

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Objective To investigate the application value of tutorial system in residents training of diagnostic ultrasound. Method 30 undergraduate residents, through the tutorial system cultivation in three years. Tutors teach their residents not only professional theory, but also systematic cultivation plan involving ward-round teaching and "hand-on" teaching practice. The teams composed of tutors and residents organize a series of teaching events such as literature review and case discussion. The role and effect of tutorial system is valued by questionnaire, test scores, and residents' subjective feelings, when they began their training system. Results 30 residency given two tests before and after the training, test results show that the accuracy after the training is significantly higher than that of before training (P < 0.05). The residents survey about satisfaction of tutorial system show that 90% residents are very satisfied, 10% are satisfied. All Residents take part in Beijing ultrasonic professional first stage examination, the results show that 100% residents pass academic knowledge test, 97% pass professional competence assessment and 93% pass reading the ultrasonography. Conclusions Tutorial system is an important part in ultrasonic resident training system, and it can cultivate qualified resident with professional theory and clinical practice ability.

Investigation of medical humanities in the perspective of clinical medicine students

2016, 36(5):  715-718. 

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Objective To learn the clinical medical students' cognition of the status quo of Chinese medical humanities and propose suggestions for improvement. Methods The survey was conducted among those medical students who have participated in clinical practice ,including undergraduates, graduate students and doctoral students . A self-filled questionnaire for all students were used for data collection． A total of 184 students completed the questionnaire．Results The survey shows that medical humanities are lack in the process of medical communication between doctors and patients. And investment in medical humanities education is insufficient .Most medical students are willing to further improve their humanistic quality through variable ways. Conclusions Medical students have a clear understanding of those problems about medical humanities and it is necessary and feasible to set up humanities related courses, organize lectures regularly, improve doctors’ salary and increase the assessment.

Parallel approach in clinical and research capacity development for medical students in the era of precision medicine

2016, 36(5):  719-722. 

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Precision medicine, as a new medical philosophy, directs the development of life science in the future, yet unknown to clinicians at large. This calls for them to correct understand, and learn how to apply in practice. As analyzed in the paper, precision medicine requires the competencies of medical students as follows: capability to take care of both clinical work and research; teamwork and leadership; as well as innovation and humanisticspirit. The authors recommend such measures as physician teacher training, multidisciplinary teamwork and teaching rounds, as well as the tutorial and co-supervisor practices. These measures can help the medical students to learn clinical abilities, and take command of the research thinking and methodology of research.