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Table of Content

    05 September 2019, Volume 39 Issue 9
    GATA-4-over-expressed bone marrow mesenchymal stem cells improve cardiac function after myocardial infarction in mice
    2019, 39(9):  1229-1233. 
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    Objective To explore the effect of GATA-4-overexpressed bone marrow mesenchymal stem cells (BMSCs) in mice to cardiac function after myocardial infarction. Methods GATA-4-overexpressed BMSCs were constructed by transfecting BMSCs with lentiviral vector carrying GATA-4. Subsequently, GATA-4-overexpressed BMSCs, GATA-4-free-vector-BMSCs, and BMSCs were subjected to hypoxia culture to induce apoptosis. After 48 h, cell apoptosis rate was determined by flow cytometry, whereas caspase-3, caspase-9, β-actin, and cytochrome C were measured by Western blot. The mouse model of myocardial infarction was used and established for 48 h. Above group of cells were injected into the tail veins. The cardiac function was examined by cardiac color Doppler ultrasonography. Moreover, the number of local apoptotic cells in myocardial infarcted mice was detected by tunnel. Results GATA-4-overexpressed-BMSCs group had stronger anti-apoptotic ability (P<0.05). The minimal expression of caspase-8 and cytochrome C occurred in GATA-4-BMSCs group (P<0.05). Ejection fraction and contraction ratio also exhibited maximal degree improvement in GATA-4-overexpressed-BMSCs group (P<0.05). Number of local apoptotic cells in myocardial infarcted mice was lower in GATA-4-overexpressed-BMSCs group (P<0.05). Conclusions GATA-4-overexpressed-BMSCs can enhance the anti-apoptotic ability of BMSCs to improve the cardiac function effectively after myocardial infarction.
    Retinal pigmentosa impairs spatial information processing of patient's peripheral visual field
    2019, 39(9):  1234-1238. 
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    Objective To investigate the spatial processing in patients with retinal pigmentosa (RP) in the central and peripheral visual fields by diffusion model analysis. Methods A total of 19 RP patients and 13 healthy controls were recruited in this study. The spatial processing task was aimed to evaluate the spatial processing in the peripheral and central visual fields with various field sizes. Reaction times (RTs), accuracies (ACCs) were recorded. In addition, the EZ-diffusion model analysis was performed for possible underlying mechanism. Results Compared with the control group [for large: (768±65) ms; for medium: (736±56) ms], RP group reacted slower in peripheral visual field including large (1193±106) ms and medium (1059±84) ms stimulus (for large: P<0.01; for medium: P<0.01). The diffusion model showed that smaller v (drift rate) (0.20±0.02) cv in large stimulus than control group (0.27±0.03) cv (P<0.05); and for large (540±44) ms, medium (556±37) ms, small (516±47) ms stimulus, the Ter (nondecision time) was found longer (for large: P<0.001, for medium: P<0.01, for small: P<0.001) than control group [for large: (286±59) ms, for medium: (358±31) ms, for small: (211±64) ms]. Conclusions RP exerts a visual field loss-related impairment in spatial cognition mainly in the peripheral visual field, which might be attributable to the decrease in information processing speed and increase in information encoding time.
    Orexin-A reduces cerebral ischemia-reperfusion injury in rats
    2019, 39(9):  1239-1242. 
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    Objective To investigate the protective effect of orexin-A (OA) on cerebral ischemia-reperfusion (I/R) injury in rats and its possible mechanism. Methods Rats were randomly divided into sham operation group (sham), ischemia-reperfusion group (I/R), 10, 30 and 50 μg/kg OA intervention group. 2,3,5-triphenylte-trazolium chloride (TTC) staining method was used to detect the cerebral infarction volume. ELISA method was used to detect the level of IL-6, TNF-α and SOD in cerebral cortex of rats, and Western blot was used to detect the expression of p-JNK. Results The I/R group had obvious infarct foci compared with the sham group, but the cerebral infarct volume significantly decreased after 30 and 50 μg/kg OA intervention (p<0.001), respectively. Compared with sham group, the levels of IL-6 and TNF-α in I / R group were significantly higher than those in sham group (p<0.05). After 30 and 50 μg/kg OA intervention, IL-6 and TNF-α levels were significantly decreased (p<0.05). The activity of SOD in I/R group was also significantly decreased (p<0.05). After 30 and 50 μg/kg OA intervention, the activity of SOD was significantly increased (p<0.05). Compared with sham group, the expression of p-JNK in I/R group was higher than that in sham group (p<0.05), yet the expression of p-JNK was decreased after 30 and 50 μg/kg OA intervention (p<0.05). Conclusion OA may play a neuroprotective role in cerebral I/R injury by inhibiting inflammatory factors and enhancing antioxidant activity.
    PODXL participates in the excessive proliferation of keloid fibroblasts
    2019, 39(9):  1243-1247. 
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    Objective To investigate the effect of Podocalyxin protein (PODXL) on the proliferation and apoptosis of keloid fibroblasts. Methods Reverse transcription quantitative PCR (RT-qPCR) was used to detect the expression of PODXL mRNA in normal skin fibroblasts and keloid fibroblasts. The expression of PODXL gene in keloid fibroblasts was inhibited by recombinant PODXL gene shRNA lentiviral expression vector infection. The proliferation of cells was measured by EdU assay. The cell cycle and apoptosis rate were analyzed by flow cytometry. The level of phosphatidylinositol 3-kinase (PI3K), tyrosine kinase (AKT) total protein and phosphorylated protein in cells were detected by Western Blot. Results Compared with normal skin fibroblasts, the expression of PODXL mRNA in keloid fibroblasts was significantly increased(P<0.05). Down-regulation of PODXL gene expression significantly inhibited the proliferation of keloid fibroblasts(P<0.05), arrested the cell cycle in S phase and promoted apoptosis(P<0.05), decreased PI3K, AKT phosphorylation protein levels(P<0.05). Conclusion The expression of PODXL in keloid fibroblasts is significantly increased. Down-regulation of PODXL gene expression may arrest cell cycle in S phase, inhibit proliferation and induce apoptosis by inhibit the activation of PI3K/AKT signaling pathway.
    Anti-inflammatory effect of metformin on LPS-induced inflammation in mice
    2019, 39(9):  1248-1251. 
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    Objective To study the anti-inflammatory effect of metformin (Met) on lipopolysaccharide (LPS)-induced inflammation in mice. Methods The mice were divided into control group, LPS group, metformin high dose group (400 mg/kg), metformin medium dose group (200 mg/kg) and metformin low dose group (100 mg/kg). The control group and LPS group were intraperitoneally injected with normal saline, the other groups were injected with different doses of metformin. After 0.5 hours, the control group was injected with normal saline, the other four groups were injected intraperitoneally with LPS. After 6 hours, the spleen index of the mice was measured. The expression of IL-6,TNF-α and IL-10 in mouse serum was detected by ELISA and the expression of IL-6,TNF-α and IL-10 mRNA in mouse liver was detected by RT-PCR. The inflammatory changes of liver tissues in each group were observed by HE staining techniques. Results Compared with the LPS group, the spleen index, IL-6 and TNF-α expression in serum and IL-6 and TNF-α mRNA expression in liver were lower in metformin group, however, IL-10 expression in serum and IL-10 mRNA expression in liver were higher in metformin group. The application of metformin(400 mg/kg) reduced the inflammatory changes of liver tissues in mouse with LPS induced inflammation. Conclusions Metformin has certain anti-inflammatory activity against inflammation induced by LPS in mice.
    Lewis lung adenocarcinoma cells regulate the phenotype transformation of macrophages in tumor microenvironment
    2019, 39(9):  1252-1258. 
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    Objective Investigating the cellular communication between Lewis lung adenocarcinoma cells (LLC) and macrophages, and exploring the possible regulatory mechanism of LLC on macrophage phenotypic transformation. Methods The normal alveolar epithelial cells (nAECs) and LLCs were co-culturing with macrophages by Transwell technique. The experiment was divided into two groups: control group and cancer cell stimulation group, in which the macrophages in lower ventricular were stimulated by LPS. After 7 days of co-culture, immunofluorescence assay was used to detect the changes of macrophage surface markers and phenotypic transformation; ELISA was used to detect the expression of inflammatory related factors IL-1β、TGF-α、IL-10 and TGF-β in the supernatant of the macrophages in two groups; Exosomes were identified by transmission electron microscopy (TEM) and Western blotting; The expression of microRNA was detected by RT-PCR in two groups. The LLC was transfected with siRNA-550a and siRNA-182 by RNAi, and then inflammatory factors expression was detected by ELISA kit. Results In the co-culture system, the expression of CD68 was high and CD163 was low in the control group (P<0.05), macrophages were M1; While in the cancer stimulation group CD68 was low and CD163 was high (P<0.05), it proved phenotype of macrophages was transformed into M2. Compared with the control group, the expression of pro-inflammatory factors IL-1β and TGF-α were high, and the expression of anti-inflammatory factors IL-10 and TGF-β were low in the cancer stimulation group (P<0.05); The results of TEM, particle size analysis and Western blotting showed there were exosomes in the supernatant of the LLC, and PKH26 staining result showed that macrophages exerted obvious phagocytic effect on exosomes. Compared with nAECs, the contents of miR-550a and miR-182 in exosomes of LLC were significantly higher (P<0.05). After transfected with siRNA-550a, the LLC exosomes significantly promoted IL-1β and TGF-α secretion of macrophages, while the levels of IL-10 and TGFβ were lower compared to siRNA-182 group (P<0.05). Conclusion LLC can transform phenotype of macrophages in tumor microenvironment into cancer supporting cells’ through secreting exosomes. The mechanism may be that MiRNA-550a in exosomes can regulate macrophages phenotype transformation.
    miR-150 inhibits cell proliferation through regulating c-Myb in human chronic myeloid leukemia cell line K562
    Lian-xiang Chen Lixia Cao
    2019, 39(9):  1259-1264. 
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    Objective To investigate the function and mechanism of miR-150 in human chronic myeloid leukemia cell line K562. Methods clinic CML patients and normal controls were collected. The peripheral blood mononuclear cells from above CML and controls were isolated. Quantitative RT-PCR analysis was used to determine the expression level of miR-150 and c-Myb mRNA; miR-150 mimic and negative control were transfected into K562 cells, respectively. CCK-8 analysis was performed to examine K562 cell proliferation. FACS analysis was used to detect cell cycle. Dual-luciferase assay and Western blot were used to detect the influence of miR-150 on target gene c-Myb. Results when compared to the normal control, the miR-150 level was down-regulated in CML patients, whereas c-Myb expression was up-regulated in CML. Overexpression of miR-150 inhibits K562 cell proliferation and cell cycle progression. miR-150 could reduce c-Myb expression in K562 cells. Conclusions miR-150 is down-regulated in CML, and it can reduce K562 cell growth by repressing the expression of oncogene c-Myb.
    miR-181 promotes the proliferation and inhibits the apoptosis of human breast cancer cell line MCF-7 via Prox1
    2019, 39(9):  1265-1269. 
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    Objective The purpose of this study is to investigate the effects of miRNA-181 and transcription homeobox factor (Prox-1) on proliferation and apoptosis of breast cancer cells. Methods The expression of miRNA-181 was detected by quantitative PCR in normal breast tissue, breast ductal carcinoma in situ tissue and breast invasive ductal carcinoma tissue. pRFP-miRNA-181-inhibitor plasmid was transfected into breast cancer cell line MCF-7, and the mRNA and protein levels of Prox1 were detected by quantitative PCR and Western-blot. Cell proliferation was evaluated by EdU assay, and apoptosis state was detected by TUNEL kit. Results Compared with normal breast tissues, the expression of miR-181 was upregulated in intraductal carcinoma and invasive ductal carcinoma (p<0.01). After the expression of miRNA-181 was inhibited in MCF-7 cells, the protein level of Prox1 was significantly upregulated (p<0.05). Morever, the proliferation ability was weakened (p<0.05) and the apoptosis ability was enhanced of MCF-7 cell (p<0.05). Conclusions miRNA-181 may play an important role in the pathogenesis of breast cancer by promoting the proliferation and inhibiting apoptosis through Prox1.
    Tracing of CM-DiI labeled BMSCs in experimental autoimmune encephalomyelitis rats
    2019, 39(9):  1270-1276. 
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    Objective: To investigate whether CM-DiI labeled BMSCs could be detected in central nervous system (CNS) of MS rat models and how they migrated. Methods BMSCs were obtained and subsequently cultured with whole bone marrow cell culture method. CM-DiI solution was used to label BMSCs in vitro and tagged cells were transplanted into EAE rats by intravenous injection through caudal vein. Tissue sections were performed and were detected under fluorescence microscope. Results CM-DiI labeled BMSCs mostly distributed around perivascular and subpial areas of spinal cord , cerebrum and cerebellum, showing round or oval shape during peak period. Cells failed to migrate into parenchyma in EAE rats. The duration is long than 2 weeks after implantation cells could be detected clearly. Conclusions CM-DiI is a good choice for BMSCs labeling and tracing. Labeled cells keep their morphology in vivo and mainly locate around vessels and subpial areas, less migrating into parenchyma of CNS.
    Decreased levels of retinol-binding protein 4 in liver tissue of hepatitis B virus infected mice
    2019, 39(9):  1277-1282. 
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    Objective To observe the effect of hepatitis B virus (HBV) infection on the expression of retinol binding protein 4 (RBP4) in mice. Methods C57BL/6 mice were randomly divided into normal group, HBV infected group(vail vein hydrodynamic injectionr AAV8-1.3HBV viral vector1×1011 vg/per)and HBV infected + lamivudine group(150 mg/kg). Then 6 mice were sacrificed at the 6 weeks, 12 weeks and 18 weeks after injection, respectively. The level of HBV DNA in serum were detected by RT-qPCR. Then, the RBP4 and biochemical indicators in mice serum were detected by ELISA and automatic biochemical analyzer; the levels of inflammatory factors(TNF-α、IL-6)in liver tissues were detected by ELISA, and the expression levels of RBP4 protein and RBP4 mRNA in liver tissue were detected by Western blotting and real-time fluorescence quantitative PCR (RT-qPCR). Results The HBV-infected mice was successfully constructed, as the serum HbsAg remained in high level and the positive rate was stable with more than 80 % at 1-18 weeks after the tail vein hydrodynamic injection, and the HBV DNA in serum of mice was also significantly higher than that in the control group. The levels of TNF-α and IL-6 in the liver tissue from the infected mice increased with time, which was significantly higher than that in the normal group at 12 weeks and 18 weeks after injection (P<0.05). In addition, the expression levels of liver RBP4 protein and RBP4 mRNA from the infected group decreased over infection time, which were lower than that in the normal group and the lamivudine group at the 6 weeks, 12 weeks and 18 weeks after injection (P<0.05). Conclusions Infection of HBV can significantly reduce the expression level of liver retinol binding protein 4 in mice.
    Urantide reduces p38 MAPK expression in thoracic aorta of rats with atherosclerosis
    2019, 39(9):  1283-1288. 
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    Objectibe To investigate the effect of urotensin antagonist, urantide on the expression of p38 mitogen activated protein kinase genes and proteins in atherosclerotic rats thoracic aorta. Methods One hundred and eighty wistar rats were randomly divided into normal group, model group, positive drug group, urantide 3d group, urantide 7d group and urantide 14d group. The model group was established by injecting a loading dose of vitamin D3 and feeding a high-fat diet. The morphological changes of thoracic aorta were detected by HE staining. The expression of p38 mitogen activated protein kinase genes and proteins situation in rat thoracic aorta was detected by immunohistochemistry, RT-qPCR and Western blot. Results Typical AS pathological changes occurred in the thoracic aorta of rats in the AS group, and urantide significantly reduced the pathological changes of the thoracic aorta. The expression of p38 MAPK and the gene and protein levels in the thoracic aorta of the AS group were significantly higher than those in the NC group (P<0.01). The positive expression of p38 MAPK and genes and proteins in the thoracic aorta of the rats in the Urantide group was significantly lower than that of the AS group (P<0.01). Conclusions Urantide can improve the function of thoracic aorta by inhibiting the expression of p38 mitogen activated protein kinase to achieve the purpose of treatment of AS.
    Role of NF2 gene in the regulation of cell cycle mediated by HCV NS4B
    2019, 39(9):  1289-1293. 
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    Objective To investigate the effect of HCV nonstructural protein 4B (NS4B) on NF2 in HEPG2 cells and the role of NF2 in NS4B regulation of cell cycle. Methods Cells were divided into several groups, one group with nothing treated and pCDNA3.1 plasmid, NS4B plasmid, NF2 plasmid, si-NF2 silencing gene and Si-control gene were transfected into other groups by cell transfection separately. RT-qPCR and Western blot were used to detect the transcription and expression of the corresponding genes. Flow cytometry was used to detect the cell cycle. Results The expression of NF2 was decreased by NS4B. The high expression of NS4B and the low expression of NF2 could both promote to S phase change in cell cycle, while high expression of NF2 arresting cell cycle in G0/G1. Conclusion NS4B can regulate NF2 and NF2 might be involved in the mechanism of regulating cell cycle by NS4B.
    Expression of HMG-CoA2-ligase in nasopharyngeal carcinoma and its influence on the cell proliferation and invasion of nasopharyngeal carcinoma cells
    2019, 39(9):  1294-1299. 
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    Objective To investigate the expression of HMG-CoA2-ligase (HMGCL) in nasopharyngeal carcinoma (NPC) and its effect on the behavior of nasopharyngeal carcinoma cells. Methods The expression of HMGCL in 64 NPCs, 42 NNE tissues, 5 NPC cell lines (HK-1, CNE1, CNE2, HONE1, HNE-1) and normal nasopharyngeal epithelial cells NP69 were detected by Western blot. Taken HK-1 cells as the research object to construct overexpressing HMGCL cells. MTT assay, Colony formation assay and Transwell assay were used to detect the proliferation, colony formation, migration and invasion of HK-1 cells after HMGCL overexpression. Results HMGCL was highly expressed in NNE and NP69 cells, and lower in NPC and NPC cell lines. HK-1 cells overexpressing HMGCL were successfully constructed. The ability of cell proliferation, colony formation, migration and invasion were significantly reduced after overexpressing HMGCL. Conclusion The low expression of HMGCL in NPCs might be related to the malignant phenotype of NPC cells. This study provides a new target for inhibiting the progression of NPC in the future.
    Prokaryotic expression and biological characteristics of recombination protein enolase – α
    2019, 39(9):  1300-1304. 
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    Objective To obtain human enolase - α( ENO1) recombinant protein by prokaryotic expression technology, and carry out bioinformatics analysis of the expressed product. Methods According to the ENO1 gene sequence (cDNA clone BC015641 MGC:23319 IMAGE:4643088) contained in GenBank, the specific primers of the gene fragment were designed. The ENO1 sequence was amplified by PCR and the clone of the gene was constructed by using the cDNA of ENO1 as a template. The vector pEASY-T1 / ENO1 and the expression vector pET-32а(+)/ENO1 were induced to express the recombinant protein ENO1 by IPTG. The bioinformatics related software (ProtParam, ProtScale, SignalP 4.1 server, Signal-3L, TMpred, DAS, NetPhos 2.0 Server) was used to analyze the physicochemical properties, signal peptides, transmembrane domains and phosphorylation sites of the recombinant protein ENO1. Results The recombinant plasmid pEASY-T1 / ENO1 and the expression vector pET-32а(+)/ENO1 were successfully constructed. The coincidence rate of the cloned plasmids and the ENO1 gene sequence in GenBank was 100%. The obtained rENO1 was about 67 ku. The ENO1 fragment consisted of 434 amino acids with a relative molecular mass of 47168.96 and a theoretical pI of 7.01, which is a stable hydrophilic protein. Conclusion Human α-dense alcoholase can be obtained by prokaryotic expression technology. The obtained fusion protein fragment is a stable hydrophilic extracellular protein, which lays a foundation for rapid early diagnosis of liver disease related markers.
    Construction and identification of luciferase reporter plasmid for human ADAM17 promoter
    2019, 39(9):  1305-1309. 
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    Objective To construct a luciferase reporter plasmid of human ADAM17 gene promoter and mutate specific sites, and to identify their biological activity. Methods FoxM1 binding sites on ADAM17 promoter were analyzed by bioinformatics. The target fragment was amplified with PCR by using genomic DNA of human gastric cancer cell line MGC-803 as template, and ligated to luciferase reporter vector pGL3-basic; the three FoxM1 binding sites were mutated on the basis of recombinant plasmid; these plasmids were transferred to 293T cells and the promoter activities were determined by double luciferase reporter gene detection system. Results The sequencing results showed that the ADAM17 promoter fluorescent reporter gene and its mutants were successfully constructed. Compared with the pGL3-basic group, the luciferase activity mediated by pGL3-ADAM17(-1 365~+51) group was significantly increased (P< 0.05); Compared with the control group, the luciferase activity mediated by overexpressing FoxM1 was significantly increased (P<0.05), and the pGL3-ADAM17(-1 365~+51) group was higher than that of the three mutation groups (P<0.05) Conclusions The human ADAM17 promoter luciferase reporter gene and its mutants are successfully constructed, and the transcriptional regulation of ADAM17 by FoxM1 is preliminarily verified.
    Interference with ARF6 inhibits proliferation, migration and invasion of human prostate cancer cell line DU145
    2019, 39(9):  1310-1315. 
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    Objective To observe the effect of interfering adenosine diphosphate ribosylation factor 6 (Arf6) on proliferation, migration and invasion of human prostate cancer cell line DU145, and to explore its regulatory mechanism. Methods DU145 cells were grown in log phase, and specific siRNA sequences and null sequences targeting Arf6 were transfected into DU145 cells by lipofection, which were set as interference group and NC group. Also take no processing as a blank group. Interference efficiency of siRNA was tested by Western blotting (Western blot). The proliferation of each group was detected and compared by MTT. The migration and invasion of each group were detected and compared by scratch test and Transwell test. Arf6, mitogen-activated extracellular signal-regulated kinase (MEK), extracellular signal-regulated kinase (ERK), Ras-associated C3 botulinum substrate 1 (Rac1) mRNA and protein expressions, phosphorylated MEK (pMEK), pERK protein expressions were detected and compared by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot. Results The interference activity of the specific siRNA sequence targeting Arf6 was (62.67±4.38)%. The cell proliferation of the interference group were significantly lower than those of the blank group and the NC group (P<0.05). The cell migration rate and the number of transmembrane cells of the interference group was significantly lower than that of the blank group and the NC group (P<0.05). The relative expressions of Arf6 and Rac1 mRNA and protein, pMEK/MEK and pERK/ERK protein in the interference group were significantly lower than those in the blank group and NC group (P<0.05). Conclusion Interfering with Arf6 can inhibit the proliferation, migration and invasion of human prostate cancer cell line DU145, which may be related to down-regulation of pMEK, pERK, Rac1 protein expression and inhibition of MEK/ERK signaling pathway.
    Value of serum RHBDD1,pepsinogen and CEA in the early diagnosis of gastric cancer
    2019, 39(9):  1316-1319. 
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    Objective To evaluate the usefulness of RHBDD1, pepsinogen(PGI and PGII) and CEA in screening atrophic gastritis, and to evaluate the value of them in early diagnosis of gastric cancer. Method We collected 78 patients from January 2016 to January 2017 who underwent gastroscopy and pathological examination of gastric mucosa in Beijing Hospital. They were divided into five groups according pathological results: CSG11cases, CAG18 cases, LGIN10 cases, EGC23 cases, AGC16 cases. Serum CEA, pepsinogen and RHBDD1 of the patients were measured. Then all the results were analysed by using SPSS19.0. Results The levels of PGⅠ/Ⅱ were statistically significant different among 5 groups (p<0.05).There were statistical differences in comparing of CSG-EGC、CSG-AGC、CAG-AGC and LGIN-AGC groups (p<0.05). Levels of CEA and PGⅠ/Ⅱ among gastritis, early gastric cancer and advanced gastric cancer were statistically different as well(p<0.05), a RHBDD1 was in ascending trend among these three groups. Conclusions Low level of PGⅠ/Ⅱ can be used in screening and early diagnosis of gastric cancer.
    Transcription factor YY1 promotes the migration ability of hypopharyngeal carcinoma cells
    2019, 39(9):  1320-1324. 
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    Objective To analyze the effect of transcription factor YY1 on the migration of FaDu cells in hypopharyngeal carcinoma. Methods Total RNA was extracted from hypopharyngeal and paracancerous normal tissues, and the expression level of YY1 was analyzed by RT- PCR. The transcription factor YY1 siRNA was transfected into the hypopharyngeal carcinoma cells, and the transfection efficiency was detected by RT-PCR and Western blot. The effect of YY1 knockdown on the migration ability of hypopharyngeal carcinoma cells was detected by Transwell experiment. Results Compared with normal adjacent tissues, YY1 was significantly upregulated in hypopharyngeal carcinoma (P<0.05). YY1 siRNA was able to reduce the relative expression of YY1 both on mRNA and protein levels significantly (P<0.05). Compared with the control group, the migration ability of hypopharyngeal carcinoma cells was significantly reduced after YY1 knockdown (P<0.05). Conclusions YY1 is a potential oncogene in hypopharyngeal carcinoma and promotes the migration of hypopharyngeal carcinoma cells in hypopharyngeal carcinoma.
    Catch-up growth of small for gestational age and its influencing factors
    2019, 39(9):  1325-1329. 
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    Objective To explore characteristics and related influencing factors of the catch-up growth of small for gestational age (SGA) infants in North China in two years after birth. Methods Newborn babies born in the Langfang Women and Children’s Health, Hebei province between January 2012 and January 2013 were enrolled in this retrospective cohort study, who were divided into SGA group, appropriate for gestational age (AGA) group and large for gestational age (LGA) group according to birth weight. Data information of infants and mothers were collected. Statistical analysis was performed to explore characteristics and related influencing factors of the catch-up growth of SGA. Results SGA 661 (8.3%), AGA 6,571 (82.5%) and LGA 729 (9.2%) were included in this study. In the SGA group, catch-up growth mainly occurred within 1 year after birth. Correlation analysis showed that the growth rate of body length within 1 year of SGA was negatively correlated with birth weight, birth body length, Apgar score and feeding patterns. It was positively correlated with the placental weight but negatively correlated with fetal/placental weight ratio (F/P). Conclusions Most SGA infants complete catch-up growth at the age of 2, and the catch-up growth mainly occurred within 1 year after birth. Inappropriately heavy placenta and feeding patterns might be related to the catch-up growth of SGA within 1 year after birth.
    Effects of a 4-hour staying at room temperature on urinary proteome analysis
    2019, 39(9):  1330-1334. 
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    Objective Using the remainder urine samples of hospital laboratories for urinary proteomic analysis to save resource for urine sample collection. Methods Urine samples collected from the same individuals were divided into two parts. In which, half of them was frozen in -80℃ immediately and the rest was stayed in room temperature for 4 h (4h-RT) before frozen in -80℃. Samples were then analyzed by proteomics strategy including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE) and liquid chromatography tanden mass spectrometry (LC-MS/MS). The differentially expressed urinary proteins were then compared between the two sample collection methods. Using the urine samples (n=150) collected by 4 h-RT, the expression levels of cytosolic non-specific dipeptidase 2 (CNDP2) between colorectal cancer (CRC) patients and health controls were analyzed by Dot blot assay. Results There was no significant difference in the main protein bands, numbers and positions of SDS-PAGE and 2DE analysis between the frozen immediately and 4 h-RT samples. A total of (1204 ± 50) kinds and (1155 ± 7) kinds of urine proteins, and (7501 ± 661) and (6940 ± 182) peptides were identified with no significant difference between the two groups. The level of CNDP2 was significantly higher in the urine of CRC patients than that in healthy controls (P < 0.001). Conclusions There was no significant difference in urinary proteome between the samples frozen immediately or 4 h-RT staying after collection. The urine samples after laboratory examinations in hospital can be used for urinary proteomics analysis. The expression level of CNDP2 is elevated in the urine of CRC patients and it may be a candidate marker of CRC.
    One case report for management of a Crohn`s disease patient with enteric stenosis and capsule endoscope incarceration by enteral nutrition
    2019, 39(9):  1337-1340. 
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    A 39-year-old male visited Peking Union Medical College Hospital with chief complain of loose stool for 2 years and abdominal pain with fiver for 2 months. He had significant loss of food intake and weight. His erythrocyte sedimentation rate (ESR) and high sensitive-C reactive protein (hs-CRP) markedly increased and Nutritional Risk Screening 2002 (NRS 2002) scored 5. CT scan revealed enteral stenosis with capsule endoscope incarceration, also entero-enteric fistula and abdominal abscess. Diagnosis of Crohn`s disease was made after excluding enteral tuberculosis. Following 4-month full-dose mesalamine and antibiotics therapy, complications didn`t improve although ESR and hs-CRP decreased evidently. Total enteral nutrition (TEN) was chosen instead of surgery. Finally, closure of fistula, disappearance of abscess, and discharge of capsule endoscopy with relief of stenosis was achieved. This case shows the promising role of enteral nutrition when handling Crohn`s disease patients with multi-complications in surgery-free situations.
    Progress on mechanism of myofibroblast activation in pulmonary fibrosis
    2019, 39(9):  1341-1345. 
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    Regardless of the etiology of pulmonary fibrosis, Myofibroblasts (MFs) play an important role in the progression of chronic inflammation and fibrosis. Persistent tissue damage is critical in initiating and perpetuating the different sources of MF precursor cells into myofibroblasts that share peculiar traits and phenotypic responses, including the ability to proliferate, migration, produce extracellular matrix components and contribute to modulation of inflammatory response and tissue angiogenesis. The activation of myofibroblast cells involves the activation of TGF -β, PDGF, CTGF and other cytokines, as well as the involvement of Wnt β-catenin signaling pathway, tissue rigidity and mechanical stretching, and microRNA.
    Progress in basic research and molecular imaging of connexin43
    2019, 39(9):  1346-1350. 
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    Connexin (Cx) is a membrane protein widely found in mammals. Its molecular weight ranges from 26Ku to 56Ku in mammals. connexin 43 (Cx43) is the most widely expressed connexin in mammals and plays an important role in the development and progression of cardiovascular diseases and tumors. In recent years, with the rapid development of molecular imaging, Cx43 as a molecular imaging target may become a new means of cardiovascular disease and tumor diagnosis and treatment.
    Exosomes in genesis, development, diagnosis and treatment of pancreatic cancer
    2019, 39(9):  1351-1355. 
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    In the microenvironment of pancreatic cancer, exosomes derived from different cell promote the growth, migration and invasion of pancreatic cancer by transferring RNA and other substances, while promoting the generation of gemcitabine resistance in pancreatic cancer. Exosomes carry a lot of information about pancreatic cancer. We can early diagnose pancreatic cancer by capturing exosomes from body fluids and analyzing their contents. Using chip technology can make detection easier and faster. One of the potential treatments for pancreatic cancer is blocking the production and transmission of exosomes or transporting antineoplastic drugs with exosomes.
    Bacteria-mediated cancer therapy by attenuated Salmonella
    2019, 39(9):  1356-1360. 
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    The use of bacteria for cancer therapy has been gradually recognized as a potential therapeutic strategy, which mainly due to the development of bacterial strains to maintain excellent anti-tumor activity and lower toxicity to host. Bacteria-mediated cancer therapy relies on facultative anaerobes that can survive in well or poorly oxygenated regions and further target tumor. Herein, Salmonella have mainly been applied as gene-delivery vectors, antitumor immune activators and tumor cell death inducers. It can ameliorate the host immune environment, reduce the expression of indoleamine-2, 3-dioxygenase(IDO) and matrix metalloproteinase 9(MMP-9) and regulate the autophagy and apoptosis of tumor cells to exert intrinsic anti-tumor activity.
    Application of next-generation sequencing technology in the connective tissue diseases
    2019, 39(9):  1361-1365. 
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    The next-generation sequencing technology (NGS) is one of the most powerful tools in modern science research with high sensitivity and specificity. Connective tissue disease is a kind of rheumatic diseases characterized by connective tissue involvement. This paper reviews the principles and the characteristics of the NGS,research profiles of connective tissue disease and its frequent diseases with a particular focus on the application in the connective tissue disease.In the end, the existing problems of NGS in life science research and future directions are prospected.
    Role of epigenetic regulation abnormality in Klotho deficiency of chronic kidney disease
    2019, 39(9):  1366-1370. 
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    Chronic kidney disease (CKD) is the persistent state of reno-protective Klotho protein deficiency. The abnormality of klotho epigenetic regulation is regarded as a major cause which reduces the expression of Klotho. Thus, it is important to elucidate the association between the abnormality of klotho epigenetic regulation and Klotho deficiency for reno-protection by targeted regulation of Klotho expression.
    Insight into the design of clinical rotation and teaching curriculum of neurology residency program at University of Massachusetts Medical School
    2019, 39(9):  1371-1375. 
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    Though China's national standardized neurology residency training has been initially established, there is still a need for improvement in the areas of design of clinical rotation and curriculum. Valuable lessons can be learned from post-graduate medical education in the United States. We reviewed relevant documents from the Accreditation Council for Graduate Medical Education (ACGME) as well as the neurology residents’ rotation schedule and teaching curriculum in the Department of Neurology at the University of Massachusetts Medical School (UMASS). Important points to note are: 1) Resident rotation schedule should meet the requirement of gradually increasing residents’ responsibilities in their training toward the goal of becoming independent practicing neurologists after their graduation. 2) Adding outpatient clinic rotations to enhance the resident learning experience. 3) In addition to daily clinical rotation and training, residency training program should design a robust curriculum based on 6 core competencies (medical knowledge; practice-based learning and improvement; interpersonal and communication skills; professionalism; and systems-based practice) to cover broad range of neurological diseases. 4) Using different lecture formats (morning report, journal club, noon conference…) to achieve different education goals.
    Questionnaire survey for evaluating gout-related knowledge for nurses
    2019, 39(9):  1376-1380. 
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    Objectives To investigate the knowledge understanding about gout for clinical nurses and improve the general nursing of gout . Methods In total, 178 clinical nurses participated in the questionnaire survey. The questionnaire has 10 items and includes the pathogenesis of gout, clinical manifestation, treatments, lifestyle and comorbidities. Results 178 clinical nurses participated in the questionnaire survey and 177 valid questionnaires were collected. All nurses correctly answered 6.96±1.67 items on average, the median was 7. The nurses were considered to acquire the gout-related knowledge if they correctly answered 7 or more items. The nurses had good understanding about the clinical manifestations of gout, but didn’t have sufficient knowledge about the treatments of gout. Logistic regression analysis revealed that further education program acted as a factor affecting the degree of acquiring the gout-related knowledge by nurses. Conclusions The gout-related knowledge of clinical nurses may be strengthened by the further education program.