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2017 Vol. 47, No. 2 Published: 2017-02-25   79 Establishment and its impact evaluation of a gastric cancer cell line with inducible expression of human cytomegalovirus UL138 protein ZHANG Liang, CHEN Wenjing, GUO Gangqiang, SUN Xiangwei, YE Lulu, HU Changyuan, JIN Jinji, SHEN Xian, XUE Xiangyang. Objective: To establish a stably inherited gastric cell line with inducible expression of human cytomegalovirus UL138 protein and then to evaluate the biological effect of UL138 protein on gastric cancers (GCs), which will offer a convincing tool for researching the specific function of UL138 protein impact on GCs. Methods: Transfect pCMV-tet3G plasmids into BGC-823 and then screen for stablely transfected clones BGCTet-on by G418 and luciferase assay. Then transfect the recombinant plasmid pTRE3G-UL138 into the selected clone and screen for stably transfected clones BGC-UL138Tet-on by G418 and hygromycin. Validate the expression of UL138 induced by doxycycline (DOX) by western blot. Besides, detect the growth impact of UL138 protein on GCs by flow cytometry and CCK8 test. Establish GC xenograft nude mice models and verify the GC inhibition function of UL138 protein in vivo. Results: Gastric cancer cell line BGC-UL138Tet-on with DOX inducible expression of human cytomegalovirus UL138 protein was successfully established. Expression of UL138 protein inhibited proliferation of GC cells distinctly. Flow cytometry test indicated UL138 protein could block GC cell cycle at G1 phase. The data of GC xenograft nude mice models further demonstrated that the BGC-UL138Tet-on xenografts were decreased, and even disappeared, when UL138 expression was induced by DOX intraperitoneal injection. Conclusion: We successfully established a specific GC cell line with inducible expression of human cytomegalovirus UL138 protein and then we found HCMV UL138 gene could inhibit proliferation of GC cells in vitro and in vivo and block its cell cycle. 2017 Vol. 47 (2): 79-84 [Abstract] ( 736 ) HTML (1 KB)  PDF (1837 KB)  ( 943 ) 85 Curcumin inhibits neuroinflammation mediated by amyloid beta-protein in BV2 cells WANG Xiaoqing, WANG Zhongsu, ZHANG Ying, CAO Hong, LI Jun. Objective: To explore the relationship between high mobility group box1 (HMGB1) and inflammatory response of the receptor for advanced glycation end product (RAGE) with inflammation model of Alzheimer’s disease (AD) selected Aβ25-35-induced BV2 cells for 24 hours later, to further investigate the effects of curcumin on expression of HMGB1, RAGE, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) in Aβ25-35-induced BV2 cells. Methods: Cultured BV2 cells in logarithmic growth phase were divided into 4 groups: control group (group A, non-treament), model Aβ25-35 group (group B, 40 µmol/L Aβ25-35, 24 h), Aβ25-35+anti-RAGE antibody group (group C, 10 µmol/L anti-RAGE antibody 1 h before+40 µmol/L Aβ25-35, 24 h) and Aβ25-35+curcumin treatment group (group D, 8 µmol/L curcumin 1 h before+40 µmol/L Aβ25-35, 24 h). The morphological character of BV2 cells was observed 24 hours later and cells viability was examined by CCK8, the level expression of HMGB1, NF-κB, RAGE in cells were detected by western blotting. The level secretion of HMGB1, IL-1β, TNF-α were detected by ELISA 24 hours later. Results: Compared with group A, the cell viability in group B and C were significantly declined and the level of HMGB1 protein expression in cells was significantly increased (P<0.05), the expression of total NF-κB were significantly increased (P<0.05), IL-1β and TNF-α in supernatant were significantly increased (P<0.05). Compared with group B and C, the cell viability, the level of HMGB1 and NF-κB protein expression in cells significantly declined, HMGB1/IL-1β and TNF-α in supernatant significantly declined in group D (P<0.05). Conclusion: Curcumin may reduce Aβ25-35-induced neuroinflammation in BV2 cells through inhibiting HMGB1 expression/extracellular released and inhibition NF-κB pathway, partly correlated with RAGE expression down-regulatd. 2017 Vol. 47 (2): 85-89 [Abstract] ( 641 ) HTML (1 KB)  PDF (1347 KB)  ( 1163 ) 90 Study on frontal lobe and occipital lobe metabolism in different stages of diabetes using 1H NMR spectroscopy WANG Dan, ZHENG Yongquan, ZHAO Liangcai, ZHENG Hong, GAO Hongchang, ZHANG Huajie.  To investigate the metabolic changes of frontal lobe and occipital lobe in different stages of diabetes. Methods: The frontal lobe and occipital lobe were dissected from the 1-week, 5-week and 9-week diabetic rats and the age-matched controls. 1H NMR-based metabonomics combined with multivariate pattern recognition analysis and quantitative analysis were used to study the metabolic characteristics. Results: The metabolic patterns of frontal lobe and occipital lobe were significantly changed in different stages of diabetes. Compared to the controls, lactate, taurine and myo-inositol were distinctly increased both in frontal lobe and occipital lobe with the development of diabetes, while the levels of N-acetyl-aspartate, aspartate and succinate were markedly decreased. Conclusion: The metabolic changes in frontal lobe and occipital lobe are similar, and occurr in the early stage of diabetes. The metabolic changes mainly focus around the disordered energy metabolism, inhibit Glu-Gln-GABA cycle and increas osmoregulation. 2017 Vol. 47 (2): 90-95 [Abstract] ( 715 ) HTML (1 KB)  PDF (1928 KB)  ( 950 ) 96 Applications of a dual-input pharmacokinetic model of dynamic contrast-enhanced MRI for evaluating tumorous microvascular properties in advanced hepatocellular carcinoma ZHAO Zhigang, ZHAO Zhenhua, YANG Jianfeng, ZHANG Yu, ZHAO Li, YANG Liming, WANG Ting, LU Zengxin. Objective: To investigate the feasibility of a dual-input two-compartment tracer kinetic model for evaluating tumorous microvascular properties in advanced hepatocellular carcinoma (HCC). Methods: From January 2014 to April 2015, pharmacokinetic parameters included transfer constant (Ktrans), plasma flow (Fp), permeability surface area product (PS), efflux rate constant (Kep), extravascular extracellular space volume ratio (ve), blood plasma volume ratio (vp), and hepatic perfusion index (HPI) were prospectively measured and analyzed using dual-input two-compartment tracer kinetic models included a dual-input extended Tofts model and a dual-input 2-compartment exchange model (2CXM) in 28 consecutive HCC patients. A paired Student’s t-test and a nonparametric paired Wilcoxon rank sum test were used to compare the equivalent pharmacokinetic parameters derived from the two models, and pearson correlation analysis was applied to observe the correlations among all equivalent parameters. The tumor size and pharmacokinetic parameters were tested by pearson correlation analysis, while correlations among stage, tumor size and all pharmacokinetic parameters were assessed by spearman correlation analysis. Results: The Fp value was greater than the PS value (Fp=1.07 mL/(mL•min), PS= 0.19 mL/(mL•min) in the dual-input 2CXM. HPI was 0.66 and 0.63 in the dual-input extended Tofts model and the dual-input 2CXM, respectively. There were no significant differences in the Kep, Vp, and HPI between the dual-input extended Tofts model and the dual-input 2CXM (P=0.524, P=0.569, P=0.622, respectively). Except for Ve, other equivalent pharmacokinetic parameters were correlated in the two dual-input two-compartment pharmacokinetic models; both Fp and PS in the dual-input 2CXM were correlated with Ktrans derived from the dual-input extended Tofts model (r=0.566, P=0.002; r=0.570, P=0.002); Kep, Vp, and HPI between the two kinetic models were positive correlated (r=0.594, P=0.001; r=0.686, P=0.0001; r=0.391, P=0.004, respectively). In the dual input extended Tofts model, Ve was significantly less than in the dual input 2CXM (P=0.004), and no significant correlation was seen between the two tracer kinetic models (r=0.276, P=0.156). Neither tumor size nor tumor stage was significantly correlated with any of the pharmacokinetic parameters obtained from the two models (P>0.05). Conclusion: A dual-input two-compartment pharmacokinetic model can be used in assessing the microvascular physiopathological properties in the pretreatment of advanced HCC. The dual-input extended Tofts model may be more stable in measuring the Ve. However, the dual-input 2CXM may be more detailed and accurate in measuring microvascular permeability. 2017 Vol. 47 (2): 96-103 [Abstract] ( 719 ) HTML (1 KB)  PDF (1798 KB)  ( 932 ) 104 Comparison of the effect of traditional Chinese medicine and Voluven on preventing ovarian hyper-stimulation syndrome during in vitro fertilization and embryo transfer WANG Peiyu, ZHAO Junzhao, LIN Jia, LU Yibin, ZHENG Jiujia, JIN Congcong, JIN Wumin Objective: To compare the effect and clinical outcome of traditional Chinese medicine and Voluven on preventing ovarian hyper-stimulation syndrome during in vitro fertilization or intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods: Prospective randomized case-control study was conducted. 74 women with high risk of OHSS on human chorionic gonadotropin (HCG) injection day were divided into 3 groups, 24 in traditional Chinese medicine group (Group A), 24 in Voluven group (Group B) and 26 in the combined group (Group C). In Group A, traditional Chinese medicine was used for 7 days since ovum pick up (OPU) day, in Group B, 500 mL Voluven vein injection were used for three days, and in Group C, traditional Chinese medicine and Voluven were both used. We compared their laboratory and clinical outcomes, vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and Nitric oxide (NO) in both serum and follicular fluid on HCG injection day, OPU day, and ET day. Results: The pregnant rate of Group A (72.22%) was higher than that of Group B (52.63%) and Group  (58.82%), but without statistic differences. No patients developed severe OHSS. There were no statistically differences in the moderate and mild OHSS rate and the result of VEGF, IL-6 and NO in all three groups. Conclusion: Both traditional Chinese medicine and Voluven have influence on the concentration of VEGF, IL-6 and NO and can avoid severe OHSS. The use of Voluven seems to produce a higher pregnant rate. Combined method can not improve the clinical and laboratory result. 2017 Vol. 47 (2): 104-109 [Abstract] ( 698 ) HTML (1 KB)  PDF (1511 KB)  ( 1061 ) 110 Expression and activity identification of Plasmodium falciparum histone H3 methyltransferase SET7 catalytic domain ZHANG Liangliang, CAI Liya, WEI Qimei, JIANG Lubin, LIANG Shaohui. Objective: To construct the recombinant baculovirus expression plasmid pFast-N-set7cd, pFast-C-set7cd and cell-free wheat germ expression vector pEU-His-set7cd, to express Plasmodium falciparum histone-lysine N-methyltransferase SET7 catalytic domain (PfSET7cd), and to identify PfSET7cd protein methyltransferase activity. Methods: pFast-N/C-set7cd and pEU-His-set7cd were constructed for expression of recombinant PfSET7cd by baculovirus-insect cell system and wheat germ cell-free expression system, respectively, and recombinant protein was identified by SDS-PAGE and Western blotting. Furthermore, recombinant protein was purified and applied in vitro enzymatic activity assay. Results: The recombinant plasmid pFast-N-set7cd and pFast-C-set7cd were constructed successfully and conformed by sequencing. However, no soluble PfSET7cd protein was detected by Western blotting in insect cells. By wheat germ cell-free expression system, soluble recombinant PfSET7cd was expressed and the molecular weight of protein agreed well with the theoretical prediction, which was identified by Western blot. And then, recombinant PfSET7cd was purified with NTA-Ni2+ affinity column. In vitro enzymatic activity assay showed that PfSET7cd exhibites H3K36me3 activity. Conclusion: Soluble recombinant protein, PfSET7cd, can be successfully obtained by wheat germ cell-free expression system, and this protein can catalyze H3K36 trimethylation (H3K36me3) in vitro while has no effects on the H3K4 trimethylation (H3K4me3) or H3K9 trimethylation (H3K9me3). 2017 Vol. 47 (2): 110-114 [Abstract] ( 655 ) HTML (1 KB)  PDF (1419 KB)  ( 738 ) 115 Expression of fibroblast activation alpha protein in glioma and its significance CHEN Yuan, SU Zhipeng, JIN Yong, ZHENG Weiming. Objective: To explore the expression of FAP-α in human gliomas, and its relationship between clinical characteristics and prognosis. Methods: Expression level of FAP-α protein was detected with immunohistochemistry in 33 cases of human gliomas and 7 cases of normal brain tissue, followed by subsequent clinicopathological correlative studies. Meanwhile, all patients were followed up, the relationship between the expression level of FAP-α and the glioma prognosis were studied. Furthermore, brain tumor molecular database (Rembrandt) was searched to investigate the relationship between the FAPmRNA expression and prognosis. Results: Comparing the expression intensities (IOD) of FAP-α protein in low-grade gliomas with those in high-grade gliomas, it was significantly different (P<0.01); the expressions of FAP-α and the pathological grades of glioma were positively correlated (r=0.673, P<0.01). There was low or no expression in 7 cases of normal brain tissue; Comparing with low grade glioma (P<0.05) and high grade glioma (P<0.01), there was significantly difference. The FAP-α expression was not significantly correlated with gender or tumor size or surgical degree and age (P>0.05). By survival analysis, overall survival time was significantly difference between the Low FAP-α and High FAP-α (P<0.01), and the expression of FAP-α was a independent prognostic factor in human gliomas (P<0.05). According to the results obtained from Rembrandt, patients with high FAP mRNA-expressing glioma showed statistically poorer prognoses compared with patients with low FAP mRNA-expressing glioma. Conclusion: FAP-α is expressed in human gliomas, which is positively correlated with the pathological grade of gliomas, and its high expression may suggest a poor prognosis. 2017 Vol. 47 (2): 115-118 [Abstract] ( 793 ) HTML (1 KB)  PDF (1683 KB)  ( 997 ) 119 Research on combination of methylphenidate and ω-3 polyunsaturated fatty acid in children with attention deficit hyperactivity disorder JIN Weimin, HUANG Yumao, YANG Xiulu, JIN Guoxin. Objective: To explore the effect of combination of methylphenidate and ω-3 polyunsaturated fatty acid (PUFA) in children with attention deficit hyperactivity disorder (ADHD). Methods: Fifty ADHD patients according with the conditions were randomly divided into two groups: group A and group B. Cross-design of two stages (I, II) was used during observation. Group A received methylphenidate and ω-3 PUFA as an intervention group for ahead of 3 months (I stage), while group B as a control group (methylphenidate and placebo). Group B was given ω-3PUFA and methylphenidate in late 3 months (II stage), while Group A as control group. Chinese version of Swanson Nolan and Pelham, Version IV Scale—parent form (SNAP-IV) were used to assess the treatment effect at the beginning and end of each stage. Adverse events were recorded. Results: The two groups both showed a reduction of attention deficit score and hyperactive/impulsive score after receiving methylphenidate and ω-3PUFA intervention (P<0.05), while oppositional defiant score showed no significant difference (P>0.05). Two-period crossover design analysis of variance showed that there was significant difference effect on attention deficit score and hyperactive/impulsive score between combination of methylphenidate and ω-3PUFA and combination of methylphenidate and placebo (P<0.05). In both two phases, the incidence of adverse events had no significant difference between the intervention group and the control group (P>0.05). Conclusion: ω-3PUFA is a safe and effective adjuvant treatment for ADHD children. 2017 Vol. 47 (2): 119-123 [Abstract] ( 749 ) HTML (1 KB)  PDF (1277 KB)  ( 959 ) 124 The effect of epinephrine on reversing asystole induced by different concentrations of bupivacaine in isolated rat hearts CHEN Hongfei, CAI Xixi, XIA Fangfang, CHEN Limei, WANG Quanguang, XU Xuzhong. Objective: To determine the effect of epinephrine on reversing asystole induced by different bupivacaine concentrations in isolated rat hearts. Methods: Thirty-six male Sprague-Dawley rats were randomly divided into six groups (n=6), then the isolated hearts asystole model induced by bupivacaine were constructed. The control group was perfused with 0.15 μg/mL epinephrine; B10, B20, B30, B40 and B50 groups were perfused with 10, 20, 30, 40 or 50 µmol/L bupivacaine and 0.15 μg/mL epinephrine respectively. The values of heart rate (HR) and rate-pressure product (RPP) were recorded after recovery of heartbeat. Results: Isolated hearts in each group achieved recovery of heartbeat but the B50 group. The rank order of HR and RPP during 15 minutes after recovery of heartbeat were the control group>the B10 group>the B20 group>the B30 group>the B40 group (P<0.01). The ratio of the maximum RPP during recovery to baseline value (y) and the concentrations of bupivacaine (x) showed a linear relationship. The linear regression equation was y=-2.385x+114.5, pearson correlation coefficient r=-0.993. Conclusion: In the isolated heart level, the resuscitation effect of epinephrine and the concentration of bupivacaine are negatively correlated. While the bupivacaine concentration is 50 µmol/L, the resuscitation effect of epinephrine has been limited. 2017 Vol. 47 (2): 124-127 [Abstract] ( 717 ) HTML (1 KB)  PDF (1310 KB)  ( 951 ) 128 The diagnostic value of contrast enhanced ultrasound in thyroid nodules with an initial inconclusive diagnostic fine needle aspiration CHEN Jianshe, ZHANG Chao, LUO Jieli, SUN Yang, HUANG Pintong. Objective: To evaluate the diagnostic value of the contrast enhanced ultrasound (CEUS) in the thyroid nodule less than 0.5 cm with an initial inconclusive diagnostic fine needle aspiration cytology (FNA).Methods: A retrospective analysis of the clinical data of 61 patients was performed, including 63 thyroid nodules with L<0.5 cm. All the thyroid nodules had an initial inconclusive diagnostic FNA and undergone CEUS, a repeat FNA after 3 months and finally confirmed by pathology. All the imaging data of preoperative FNAs and CEUS were collected. Pursuant to which, the detection rate of malignant nodules (DROMN), the detection rate of begin nodules (DROBN), the false negative rate (FNR), the false positive rate (FPR), the accuracy of diagnosis, the persistently inconclusive diagnostic rate (PIDR) and the malignant proportion by repeat FNA and CEUS were analyzed respectively. Results: In CEUS group, the DROMN, DROBN, FNR, FPR, the accuracy of diagnosis, PIDR and the malignant rate were 84.8%, 36.7%, 15.3%, 12.5%, 86.7%, 28.5% and 16.7% respectively. In the repeat FNA group, DROMN, DROBN, FNR, FPR, the accuracy of diagnosis, PIDR and the malignant proportion were 24.2%, 33.3%, 41.2%, 0.0%, 72.0%, 60.3%, 47.4%. The DROMN, PIDR and the malignant rate in the group CEUS were statistically higher than the repeat FNA group (P<0.05). Conclusion: CEUS has a better diagnostic efficiency in the thyroid nodule less than 0.5 cm with an initial inconclusive diagnostic FNA than the repeat FNA. The non-invasive CEUS examination can be suggested. 2017 Vol. 47 (2): 128-131 [Abstract] ( 642 ) HTML (1 KB)  PDF (1441 KB)  ( 841 ) 132 Comparison of late preterm infants and full term in the early growth and development WANG Qingqing, SU Weidong, HUANG Yudan, ZHANG Weiwei. Objective: To evaluate physical, neurological development and prognosis through one year follow-up of late preterm infants. Methods: Choose our hospital obstetrics between April 2013 and March 2013 in our hospital was born and in the late outpatient follow-up premature as observation object (n=50), randomly selected the born healthy full term for the same period as the control group (n=60). Late preterm infants at 0 month (40 weeks calibration), 6 and 12 months to mass, length, head circumference measurement, compared with the same age full term. Between 0 and 1 months late preterm neonatal neurobehavioral determination (NBNA) scores, and the determination results compared with the full term of on the same day age. Two groups at follow-up to correction in 6 and 12 months of age: Gesell examination, early intelligence development level. Results: Late preterm group’s body length and head circumference were less than the full term on the 0 and 1 month, the difference was statistically significant (P<0.05). Late preterm group at 0 and 1 months, NBNA scores were lower than the full term on the same age group, differences were statistically significant (P<0.05). Six months late preterm big movement lagged behind the full term, 12 months when the big movement, fine motor, language behind the full term differences were statistically significant (P<0.05). Conlusion: Compared with the full term infants, born in late preterm birth after early physical and neurological development lags behind that of the full term, to strengthen monitoring and targeted intervention. 2017 Vol. 47 (2): 132-135 [Abstract] ( 774 ) HTML (1 KB)  PDF (1249 KB)  ( 1184 ) 136 Application of multidisciplinary team pattern in diagnosis for the small pulmonary nodules YAN Boer, ZHANG Yongkui, LE Hanbo, CHEN Zhijun, ZHANG Binjie, WANG Shanjun Objective: To evaluate the effect of multidisciplinary team (MDT) pattern applied in diagnosis for small pulmonary nodules. Methods: We retrospectively analyzed changes in the amount of the outpatients for small pulmonary nodules, the cases of detected small-size lung cancer, the rate of diagnostic agreement of small pulmonary nodules and the quantity of surgery for small-size lung cancer after application of the MDT pattern. Results: The number of the outpatients for small pulmonary nodules increased from 1 136 patients in 2012 to 4 455 patients in 2014, and the cases of the detected small-size lung cancer increased from 56 in 2012 to 118 in 2014 with tumor diameter≤1 cm, and from 106 to 210 with diameter≤2 cm, respectively, and the rate of diagnostic agreement raised from 83.4% in 2012 to 91.4% in 2014 after the MDT pattern was applied. Conclusion: Application of the MDT pattern significantly increases the outpatients for small pulmonary nodules. Besides, the small-size lung cancers are detected more efficiently, and the diagnostic accordance rate is accordingly improved. 2017 Vol. 47 (2): 136-139 [Abstract] ( 679 ) HTML (1 KB)  PDF (1310 KB)  ( 961 ) 140 Expression of dipeptidyl peptidase IV gene in gastric carcinoma and its’ signifance for prognosis TIAN Jie, ZHANG Zhi’an, SUN Guoxing, XIAO Baozeng, Zhang Wenchao, Yang Jitao, Zhang Qingyong, Yin Xiao, Geng Guowang. Objective: To evaluate the expression of dipeptidyl peptidase IV (DPP IV) in gastric carcinoma tissue and matched para-tumor tissues, then its clinical pathologic significance for gastric carcinoma was investigated. Methods: One hundred and twelve cases gastric carcinomas tissues and the same number of matched para-tumor tissues were selected, expressive difference of DPP IV gene between carcinoma tissues and their matched para-tumor tissues was measured by immunohistiochemistry; with protein concentration level of DPP IV measured by Western blotting methods. Results: The expressive loss number for DPP IV was 3 (2.7%) in 110 normal gastric tissues, while lossed in 78 gastric carcinomas; and the level of DPP IV decreased obviously in DPP IV positive tissues; DPP IV is an independent prognostic factor of gastric carcinoma by multiple factor analysis. Conclusion: Loss of DPP IV’s expression is an independent poor prognostic factor of gastric carcinoma. 2017 Vol. 47 (2): 140-143 [Abstract] ( 768 ) HTML (1 KB)  PDF (1418 KB)  ( 801 )

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