Journal of Shandong University (Health Sciences)

Effects of puromycin-sensitive aminopeptidase on the apoptosis induced by TauP301L in the cultured cells

WANG Miaomiao1, DIAO Xueqin2, ZHUANG Yunlong3, XU Gang4, ZHOU Nan1, JIA Yanglei1, LI Mengmeng1, REN Guijie1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 1-7. doi:10.6040/j.issn.1671-7554.0.2013.618

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Objective To explore the effect of puromycin-sensitive aminopeptidase (PSA) on the apoptosis induced by TauP301L in SH-SY5Y cells and PC12 cells. Methods The proliferation and morphological changes of the cells were detected after SH-SY5Y and PC12 cells were transfected with TauP301L for 24, 48 and 72h, respectively. The optimal time for TauP301L transfection was determined. To evaluate the effect of PSA on cell apoptosis, PC12 cells were transfected with PSA and TauP301L, and SH-SY5Y cells were transfected with PSA-siRNA and TauP301L. Cell apoptosis was determined by flow cytometry. The expression and activity of Caspase-3 were detected by Western blotting and Microplate reader, respectively. Results TauP301L induced cells apoptosis in a time-dependent manner. After TauP301L transfection for 48h, the proliferations of SH-SY5Y and PC12 cells significantly decreased (P<0.01), while the apoptosis of the cells increased (P<0.01),as well as apoptosis-like cells could be found after Hoechst33342 staining of cell nuclei. Compared with TauP301L transfection, the expression and activity of Caspase-3 were downregulated (P<0.01), and the apoptosis decreased (P<0.01) after transfection with both PSA and TauP301L in PC12 cells. However, the expression and activity of Caspase-3 were upregulated (P<0.01) and the apoptosis increased (P<0.05) after transfection with both PSA-siRNA and TauP301L in SH-SY5Y cells. Conclusion TauP301L can induce apoptosis in both SH-SY5Y cells and PC12 cells. PSA has a protective effect on nerve cells by inhibiting apoptosis induced by TauP301L.

Role of tumor-associated macrophages in epithelial-mesenchymal transition of human hepatocellular carcinoma cells

WANG Hao1, LI Xia2, WANG Chao3, LI Guosheng1, GUO Chun1, ZHU Faliang1, ZHANG Lining1, SHI Yongyu1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 8-12. doi:10.6040/j.issn.1671-7554.0.2013.554

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Objective To explore the effect and mechanism of tumor-associated macrophages on human hepatocellular carcinoma (HCC) cells. Methods The HCC cells were cocultured with macrophages from PMA-treated THP-1 cells and cell migration was detected by transwell migration test and wound-healing assay. The expressions of E-cadherin and N-cadherin were measured by RT-PCR. Results The ability of cell migration was significantly increased after being cocultured with mocrophages from PMA-treated THP-1 cells. A transition from epithelial morphology to mesenchymal morphology was observed. The expression of E-cadherin decreased and the expression of N-cadherin increased. Conclusion Our findings suggest that tumor-associated macrophages may promote the cell migration through epithelial-mesenchymal transition.

Effects of enhanced expression of RbAp48 mediated by recombinant adenovirus on proliferation and growth of cervical cancer cells

ZHANG Wen1, FENG Tingting1, ZHENG Lin1, WANG Hong1, LU Yi2, QI Mei1, YU Xiuping1, TANG Wei1, ZHAO Weiming1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 13-17. doi:10.6040/j.issn.1671-7554.0.2013.762

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Objective To construct the recombinant adenovirus carrying RbAp48 gene, and to study its effect on the behaviors of human cervical cancer cell line SiHa. Methods The shuttle plasmid pDC316-RbAp48 was constructed and co-transfected into the HEK293 cells with genomic plasmids pBHGlox ΔE1, 3Cre to produce the replication-deficient recombinant adenovirus Ad5-RbAp48. Then the SiHa cells were infected with Ad5-RbAp48, and RbAp48 expression was examined by immunoblotting. WST-8, cell number counting, and soft agar assay were carried out to examine the effects of Ad5-RbAp48 infection on cell proliferation and anchorage-independent growth of SiHa cells. Results Ad5-RbAp48 was successfully constructed. Ad5-RbAp48 infection significantly enhanced the expression of RbAp48 in SiHa cells, and effectively inhibited cell proliferation and anchorage-independent growth. Conclusion Recombinant adenovirus carrying RbAp48 gene can mediate RbAp48 expression and inhibit behaviors of cervical cancer cells.

Effect of PDCD4 siRNA on the lipid accumulation in macrophages#br#

DONG Zhaojing, SHANG Qianwen, GUO Chun, ZHANG Lining, WANG Qun

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 18-21. doi:10.6040/j.issn.1671-7554.0.2013.399

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Objective To investigate the effect of PDCD4 on lipid accumulation in macrophages. Methods The cells were divided into PDCD4 siRNA group and negative control siRNA group. Gene silencing was performed in murine macrophages using PDCD4 siRNA and negative control siRNA; the mRNA and protein levels of PDCD4 were detected by RT-PCR and Western blotting; after treatment with ox-LDL, the lipid contents in macrophages were detected by Oil red O staining; the expressions of cholesterol efflux-related genes liver X receptor α (LXR-α), ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) were detected by Real-Time PCR. Results PDCD4 siRNA effectively reduced the expression of PDCD4 on both mRNA and protein levels in macrophages; the lipid contents decreased significantly in PDCD4 siRNA group compared with those in negative control siRNA group (P＜0.001), and the expressions of LXR-α (P＜0.05), ABCA1 (P＜0.01) and ABCG1 (P＜0.001) were markedly up-regulated. Conclusion Silencing of PDCD4 gene can reduce the lipid accumulation in macrophages through promoting cholesterol efflux.

Rosuvastatin reverses the effect of heavy drinking on angiogenesis after infarction in rats

QU Fuchao, WANG Aihong, SUN Yongle, WANG Yong, YUAN Haitao, ZHANG Yuying

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 22-25. doi:10.6040/j.issn.1671-7554.0.2013.552

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Objective To study the effect of heavy drinking on angiogenesis in ischemic rats and the effect of Rosuvastatin on angiogenesis after heavy drinking. Methods A total of 24 male SD rats were randomly divided into 3 groups: control group, heavy-drinking group［50% ethanol, 5g/(kg·d)］ and Rosuvastatin pretreated group［Rosuvastatin (20mg/(kg·d)+50% ethanol, 5g/(kg·d)］. The three groups were given equal volume of ethanol, rosuvastatin and vehicle by gavage respectively for 4 weeks and then myocardial infarction models were constructed. The rats were killed on the 3rd day after myocardial infarction. The microvascular density (MVD) was observed by immunohistochemical SP method. The vascular endothelial growth factor (VEGF) and endostatin expressions in the ischemia myocardium tissue were tested with ELISA. Results Compared to the control group, the MVD was significantly reduced (P<0.01) in the heavy-drinking group, the level of endostatin was significantly increased (P<0.01), and there was no difference in the expression of VEGF. MVD in the Rosuvastatin pretreated group was significantly increased compared to that in the heavy-drinking group (P<0.01), but the VEGF and endostatin expressions were not significantly different between the two groups(P＞0.05). Conclusion Heavy drinking could inhabit angiogenesis by increasing the expression of endostatin after myocardial infarction. Rosuvastatin could reverse the damages of heavy drinking on angiogenesis.

Effects of vitrification on microfilaments′ distribution and development potentiality of human in vitro matured oocytes

CI Qianqian, LI Mei, WU Keliang, MA Shuiying, WANG Miaomiao, LIU Hui, GAO Qin, SHI Yuhua, CHEN Zijiang

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 26-29. doi:10.6040/j.issn.1671-7554.0.2013.619

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Objective To find the changes of development potential and microfilament structure which were affected by vitrification for human in vitro matured oocytes. Methods Human in vitro matured oocytes were gathered and randomly divided into two groups: fresh group and vitrified group. The oocytes were fertilized and immunofluorescencently stained. The numbers of 2PN, blastocyst and high quality blastocyst were calculated and the microfilaments structures were observed by laser confocal microscopy (LSCM). Results The survival rate of oocytes was 83.74% after vitrification-warming. The 2PN, blastocyst and high quality blastocyst of the fresh group were significantly higher than those of vitrified group (P<0.05). There was no statistical difference in normal microfilaments rate between fresh group and vitrified group (71.67% vs 67.50%, P=0.71). Conclusion The proper vitrification-warm process doesn’t change the microfilaments structures of human in vitro matured oocytes, but it reduces their development potential. Thus, morphology of the microfilaments alone can’t be taken as the indicator of the developmental potential.

Protective effect of autophagy on the inflammation induced by ox-LDL

HAO Fengcheng1,2, SU Zhonghua1,3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 30-34. doi:10.6040/j.issn.1671-7554.0.2013.698

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Objective To study the effect of autophagy on the inflammation induced by oxygenized low density lipoprotein (ox-LDL). Methods The THP1 cells were divided into four groups: control group, ox-LDL treatment group, 3-methyladenine (3-MA) treatment group, ox-LDL combined with 3-MA treatment group. The expression of LC3 protein was detected by Western blotting. The formation of autophagosome was measured by immunofluorescence with LC3-specific antibody. MDC staining was used to detect the intracellular acidic vesicles. The changes of inflammatory factor level was determined by ELISA analysis. Results Ox-LDL could induce autophagy in monocyte THP1. The inflammatory reaction induced by ox-LDL was significantly enhanced after inhibition of autophagy (P<0.05). Conclusion Autophagy represents a defense mechanism in the inflammation induced by ox-LDL.

Effect of hypoxia on the expressions of HIF-1α and DEC1 in human gastric cancer cell

HU Rui1, JIA Yanfei2, ZHENG Yan2, MA Xiaoli2, KONG Yi1, LI Pin1, WANG Yunshan2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 35-38. doi:10.6040/j.issn.1671-7554.0.2013.519

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Objective To investigate the effect of hypoxia on the expressions of hypoxia inducible factor-1α (HIF-1α) and differentiated embry-ochondrocyte expressed gene 1 (DEC1) in human gastric cancer cells. Methods The BGC823 cell lines were divided into the experimental groups (hypoxia for 24 hours group and hypoxia for 48 hours group), the control groups (normoxic for 24 hours group and normoxic for 48 hours group), and the negative control group (PBS instead of the first antibody). The experimental groups were cultured in the hypoxia incubator with 5%CO2 and 0.5%O2, and the control groups were exposed to the normal oxygen condition. The levels of mRNA and protein of HIF-1α and DEC1 were determined by RT-PCR, Western blotting and immunocytochemistry method, and their correlations were analyzed. Results The expressions of both mRNA and protein of DEC1 and HIF-1α in BGC823 cell lines were upregulated significantly compared with the control groups under the hypoxia condition (P<0.05). With the extension of hypoxia, the mRNA expressions of HIF-1α and DEC1 increased, while the protein expressions of HIF-1α and DEC1 declined. Immune cytochemistry method also showed that the positive expression rates of HIF-1α and DEC1 in the experimental groups were higher than those in the control groups (P<0.05). Conclusion Hypoxia can induce the expressions of HIF-1α and DEC1, which have significant correlation, in BGC823 cell lines. During hypoxia, HIF-1α protein accumulates and activates the transcription of downstream target gene DEC1, which suggests that DEC1 is a marker of activated hypoxic pathways and plays an important role in hypoxia tolerance in gastric cancer.

Effect of liraglutide on the expressions of NF-κB and VCAM-1 in aorta endothelium of type 2 diabetic rats

QI Caihui1, JIN Yongjun1, YANG Meizi2, XU Xinmiao1, ZHANG Lingyun1, CAO Pengjuan1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 39-42. doi:10.6040/j.issn.1671-7554.0.2013.589

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Objective To investigate the effect of liraglutide on the expressions of nuclear factor-kappa B (NF-κB) and vascular cell adhesion molecule 1 (VCAM-1) in aorta endothelium of type 2 diabetic rats and the possible mechanism. Methods Forty male SD rats were divided into the normal control group (NC group), diabetic control group (DM group), glibenclamide group (DMG group) and liraglutide group (DML group). The model of type2 diabetic rats was eatablised by injecting with low-dose streptozocin(STZ) into abdominal cavity of SD rats after high-fat and high-sugar diet. Then rats in DMG and DML groups were treated with liraglutide and glibenclamide for 8 weeks. At the end of the twelfth week, blood was taken from abdominal aorta to examine biochemical indexes including fasting blood-glucose(FBG), fasting insulin(FIns), totalcholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C) and high density lipoprotein-cholesterol (HDL-C). The thoracic aorta was isolated to observe the histological parameters by HE staining. The expressions of NF-κB and VCAM-1 in aorta endothelium were determined by immunohistochemical method. RusultsThe levels of FIns and HOMA-IR in DMG group and DML group were decreased compared with those in DM group (P<0.01). The levels of FIns, HOMA-IR,TG and LDL-C and the expressions of NF-κB and VCAM-1 in in aorta endothelium in DML group were decreased compared with those in DMG group (P<0.05). Conclusion Liraglutide can decrease the expressions of NF-κB and VCAM-1 in aorta endothelium of type2 diabetic rats and improve glucolipid metabolism. This may be involved in vascular complications of diabetes.

Effect of SIRT3 siRNA on ICAM-1 and E-selectin expressions in HAEC stimulated by ox-LDL

DING Ling1,2, GONG Huiping1, DU Yimeng1, WANG Xin1, ZHANG Yan3, WANG Xuping2, LU Qinghua1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 43-48. doi:10.6040/j.issn.1671-7554.0.2013.532

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Objective To investigate the effect of SIRT3 gene on the expressions of inflammatory factors intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin in human aortic endothelial cell (HAEC) stimulated by oxidized-low density lipoprotein(ox-LDL). Methods The cells were randomly divided into normal control group, transfection reagent control group, negative control group, and SIRT3 siRNA (1, 2, 3 and 4) groups. When the most effective interference sequence was selected out, the cells were divided into normal control group, siRNA group, (20, 30 and 40μg/mL) ox-LDL groups and siRNA+(20, 30 and 40μg/mL) ox-LDL groups. The cells were transfected for 24h and stimulated by ox-LDL for 6h. SIRT3 mRNA level was detected by qRT-PCR and its protein level was analyzed by Western blotting. Mean fluorescence intensities (MFIs) of ICAM-1 and E-selectin antibody were detected by flow cytometry. Results SIRT3siRNA2 was the most effective interference sequence. The interference efficiency of SIRT3siRNA2 was 88.62% in mRNA level and 80.37% in protein level. Expressions of SIRT3 mRNA had significant differences in ox-LDL groups (all P＜0.05) compared with that in normal control group; with the increase of ox-LDL concentration, the expression of SIRT3 mRNA gradually declined (all P＜0.05). MFI of ICAM-1 and E-selectin had statistical differences in siRNA+ox-LDL groups compared with that in siRNA group (all P＜0.05); with the increase of ox-LDL concentration, the MFI was enhanced obviously (all P＜0.05). MFI of ICAM-1 and E-selectin in siRNA group had no significant difference compared with that in normal control group (P＞0.05). Conclusion The SIRT3 gene has association with the expressive increments of E-selectin and ICAM-1 in HAEC stimulated by ox-LDL, and may regulate the inflammatory reaction in artery endothelial cell injury process.

Establishment of cisplatin-resistant gastric cancer cell line and the biological characteristics

YAN Bing1, LIU Guoqin2, JIANG Yongsheng2, SHEN Haiyu2, SUN Shaochuan2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 49-52. doi:10.6040/j.issn.1671-7554.0.2013.556

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Objective To explore the drug-resistant gastric cancer cell line induced by high-dose cDDP and its biological characteristics. Methods A drug-resistant gastric cancer cell line BGC823/cDDP was established by high-dose cDDP. The changes of biological characteristics of the cell lines were observed using light microscopy. The resistance index was determined by CCK-8 assay. The cell growth curve was drawn by CCK-8 assay. The cell cycle of drug-resistant cells was tested by flow cytometry. The cell invasion ability was observed by Transwell assay. The densities of CEA and CA-199 in the cell culture medium were evaluated by immune luminescence. Results Compared with BGC823 cells, the BGC823/cDDP cells turned to be slightly bigger and irregular in shape, and some giant cells could be seen. The proliferation rate of BGC823/cDDP cell prolonged, with the resistance index being 4.54. The population doubling-time prolonged. The cell numbers of G1 phase increased while decreased in S and G2 phase in BGC823/cDDP cell. The invasion ability was stronger in BGC823/cDDP cell (P<0.05). The densities of CEA and CA-199 had no significant differences between BGC823 and BGC823/cDDP. Conclusion The cDDP-resistant gastric cancer cell line BGC823/cDDP was induced by high-dose cDDP. The BGC-823 and BGC-823/cDDP cells were different in biological characteristics.

Initial exploration on the recycling of deep second-degree burn skin for constructing model of complex skin transplanting

ZHU Xuguo1,2,3, YU Guanying3, LI Yaonan1,2,3, LIU Lei1, ZHANG Jixun1, LIU Zhenzhong1, JIANG Duyin1,2,3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 53-57. doi:10.6040/j.issn.1671-7554.0.2013.615

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Objective To explore the feasibility of deep second-degree burn skin (DBS) as dermal substitute for constructing model of complex skin transplanting in repairing wounds. Methods A total of 20 Wistar rats were randomly divided into the deep second-degree burn dermal matrix (DBDM) transplanting group (experimental group) and acellular dermal matrix (ADM) transplanting group (control group), 10 rats in each group. After a second-degree scald, the skin of two rats in experimental group was treated with enzyme digestion to remove cells to prepare DBDM. The back skin of newborn rats was integrally taken and the subcutaneous and dermal tissues were removed. The thin skin overlapped with mesh DBDM were transplanted into the back skin wound of full-thickness in size of 2cm×2cm or so. After 14 days, the situatioin of composite skin growth was observed. ADM was prepared by normal skin in control group, and other experiment steps were the same as the experimental group. Results The freshly prepared DBDM was yellow-white, less soft in texture of flexibility and extensibility compared with ADM. Collagen fibers of DBDM were thickened unevenly with hyaline changing and arranging in disorder. No cellular, blood vessels, hair follicles and other components were observed in DBDM under light microscope. Fourteen days after the composite skin transplantation experiments, the survival rate of rats and transplants in the two groups had significant difference (P<0.05). The bilayer composite skin and subcutaneous tissue connected closely. The deep DBDM or ADM was pink. The skin after operation had no significant swelling or inflammation. Infiltration of inflammatory cells and ingrowth of fibroblast and capillary appeared more earlier in DBDM group compared with those in ADM group. Inflammatory cells were mainly neutrophils and macrophages. New collagen deposition was rapid and collagen fibers arranged in a regular and dense state. Conclusion Sources of DBDM is wide and preparation method is simple. Biocompatibility of transplants is good and burn toxin-like effect is weak. DBDM can act as dermal substitute of autologous split thickness skin. The experiment may provide new ideas for clinical transformation and utilization of DBS.

Association between serum testosterone and lipid profile in men with type 2 diabetes mellitus

ZHANG Nan1, YUAN Zhongshang2, ZHANG Xu1, ZHANG Yikun1, YU Chunxiao1, GAO Ling3, ZHAO Jiajun1, GUAN Qingbo1, GUAN Liying4

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 58-61. doi:10.6040/j.issn.1671-7554.0.2013.770

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Objective To evaluate the association between serum testosterone level and the lipid profile, after adjusting for some traditional confounding factors and thyroid function. Methods The total testosterone (TT), follicle stimulating hormone (FSH), luteinizing hormone (LH), total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) of 125 male patients with type 2 diabetes mellitus (T2DM) were measured. Testosterone secretion index (TSI) was calculated by dividing TT by LH. Bivariate and partial correlation analysis, and multiple linear regression analysis were performed to explore the association between TT level and lipid levels. One-way analysis of variance (ANOVA) was adopted to compare the lipid levels among four subgroups which were divided by quartile of TT (Group A: TT < 3.93ng/mL, Group B: TT 3.93-5.09ng/mL, Group C: TT 5.10-6.58ng/mL, and Group D: TT > 6.58ng/mL). Results TT was negatively correlated with TG (P<0.05), and positively correlated with HDL-C (P<0.05) while TSI was negatively correlated with TG (P<0.05) in correlation analysis and multiple linear regression analysis. Compared with the group with the lowest TT level (group A), TG level was significantly lower in groups B, C and D, and HDL-C was significantly higher in groups C and D (P<0.05). Conclusion Serum TT was negatively correlated with TG and positively correlated with HDL-C in male T2DM patients, suggesting that serum testosterone might be a protective factor for dyslipidemia in these patients.

Changes of NF-κB expression after promoter demethylation of TMS1 in K562 cells

LI Hongli, WANG Yan, XU Wenwei, DONG Lin, GUO Yan, ZHU Chuansheng, BI Kehong

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 62-66. doi:10.6040/j.issn.1671-7554.0.2013.630

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Objective To explore the association between the methylation degree of TMS1 gene in chronic myeloid leukemia cell line K562 cell and NF-κB. Methods A total of 6 samples of bone marrow of healthy controls and patients with non-hematologic malignancies were randomly collected as case-control group. Then total RNA was extracted and TMS1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). K562 cells were cultured and treated with decitabine (DCA) with different concentrations. The blank control group and experiment group (0.5μmol/L DCA, 1μmol/L DCA) were set up separately. Methylated degree of TMS1 was detected by methylation specificity polymerase chain reaction (MS-PCR), mRNA and protein of TMS1 and NF-κB in K562 cells were checked by RT-PCR and Western blot, the growth of cells was inspected by CCK-8 and morphologic changes were observed by light microscope. Results TMS1 mRNA was found in the case-control group, but was absent in K562 cells. The TMS1 gene was exhaustively methylated in the blank control group. After promoter demethylation of TMS1, the expression of NF-κB significantly decreased (P<0.05). The K562 cell proliferation was inhibited in a dose- and time-dependent manner. Conclusion TMS1 gene presents exhaustive methylation in K562 cells and its silent expression may abnormally activate NF-κB signaling pathway and plays a role in the pathogenesis of leukemia.

Siginifcance of polarizing microscope for the diagnosis of primary tracheobronchial amyloidosis and literature review #br#

YING Yangyang, MA Dedong, XIAO Wei

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 67-69. doi:10.6040/j.issn.1671-7554.0.2013.431

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Objective To explore the application of polarizing microscope in the diagnosis of tracheobronchial amyloidosis, in order to reduce the rate of misdiagnosis. Methods A total of 13 pathological sections positive for Congo red were re-observed, and 104 cases reported since 1987 which had complete clinical data and implicit pathological Results were analyzed. Results Of the 13 pathological sections, 8 showed apple-green birefringence when viewed under polarizing microscope. If the diagnosis of amyloidosis was based on Congo red staining only, it would result in 39.5% false positive cases, which might indciate that 40 cases might be misdiagnosed among the 104 confirmed cases. Conclusion Only when both Congo red staining and polarizing microscope show positive Results , can we diagnose the case as PTBA.

Plasma levels of IL-6, IL-10 and IL-12 and influencing factors in first-episode and neuroleptic-free schizophrenia patients

ZHAO Yan1, WANG Gang1, CHEN Dafang2, CHENG Yuhang1, CHEN Xueyan1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 70-73. doi:10.6040/j.issn.1671-7554.0.2013.272

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Objective To investigate the plasma levels of interleukin-6 (IL-6), interleukin-10 (IL-10) and interleukin-12(IL-12) in first-episode and neuroleptic-free schizophrenia patients and their influencing factors. Methods The 137 subjects were divided into the patient group (n=67) and control group (n=70). The plasma levels of IL-6, IL-10 and IL-12 were assayed by enzyme-linked immunosorbant assay (ELISA). The symptoms were measured by positive and negative syndrome scale (PANSS). Results Compared with the control group, the patient group had the increased IL-6 levels (P=0.028) and decreased IL-10 levels (P=0.013). No statistical difference of IL-12 levels was found between the two groups (P=0.258). There existed positive correlations between negative subscore and IL-10 levels (β=1.9, P=0.011), and the amount of cigarettes and IL-12 levels (β=0.1, P=0.049). Conclusion Schizophrenia is characterized by alteration of immunity and cytokine network damage. There may exist multi-form alterations of immunity among different subtypes and sexes of schizophrenia patients.

Role of innate immunity induced by AIM2 in the pathogenesis of chronic hepatitis B

PAN Jiachao1,2, ZHANG Le1,2, XU Linlin3, SHAN Xiaoyu2, DU Wenjun2, CHEN Shijun2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 74-79. doi:10.6040/j.issn.1671-7554.0.2013.663

Abstract ( 804 )

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Objective To explore the role of innate immunity induced by absent in melanoma 2 (AIM2) in the pathogenesis of chronic hepatitis B (CHB). Methods A total of 70 cases were enrolled in this study, including 47 CHB cases as the experimental group and 23 cases of fatty liver as the controls group. The immunohistochemical method was adopted to detect the expressions of AIM2, Caspase-1, IL-1β and IL-18 in liver tissues. The differences between the two groups were compared with chi-square test, and the correlation was analyzed using spearman test. Results The positive rate of AIM2 expression in the experimental group (89.3%) was significantly higher than that in the control group (43.5%) (χ2 = 15.655, P<0.01). In the experimental group, Caspase-1 was positively correlated with AIM2 (rs=0.738, P<0.01); IL-1β and IL-18 were positively correlated with AIM2 (rs=0.527, 0.642, P<0.01). ALT and AST were positively correlated with AIM2 (rs = 0.325, 0.362, P<0.01). The AIM2 expression in the high HBV titers group (HBV-DNA ≥ 1×105 copies/mL) was significantly higher than that in the low HBV titers group (HBV-DNA < 1×105 copies/ml）(χ2= 27.572, P<0.01). Conclusion In the innate immune response to HBV infection, AIM2 can recognize HBV-DNA, activate Caspase-1 pathways subsequently, release IL-1β and IL-18 inflammatory factors, thus leading to liver inflammatory damages in chronic hepatitis B.

Risk factors of allodynia in migraine and the therapeutic effect of pregabalin

YU Feiyu, ZHANG Na, CHEN Chunfu

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 80-84. doi:10.6040/j.issn.1671-7554.0.2013.584

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Objective To observe the predictors of cutaneous allodynia（CA） in migraine patients and the effects of pregabalin in alleviating CA. Methods Sixty-three patients with migraine were included and divided into two groups，CA group（41 patients）and non-CA group（22 patients），according to Allodynia Symptom Checklist（ASC）. The presence of CA, its type and severity, and details of headache were enquired and the predictors of CA were analyzed. Forty-one patients with CA were treated with pregabalin (150-300 mg/d) for 12 weeks. The improvements of allodynia and headache were recorded after 1 and 3 months treatment and the effect of pregabalin on CA was discussed. Side effects of the drug were recorded. Results CA appeared in 65.1% patients with migraine， including thermal CA in 85.4%， static mechanical CA in 29.4% and dynamic CA in 22.0%. CA was associated with gender, duration of illness and medication-overuse. Compared with baseline, pregabalin significantly reduced the severity and the number of patients with allodynia after 1 and 3 months treatment (P<0.05). The frequency and severity of headache, MIDAS and HIT-6 scores significantly decreased after the treatment of pregabalin for 3 months compared with baseline(P<0.05). The total therapeutic effect of pregabalin was 85.4%. Conclusion CA appears in 65.1% patients with migraine. It is associated with gender, duration of illness and medication-overuse. Pregabalin may be effective in relieving allodynia in migraine.

Clinical implication of psychotherapy on elderly patients with coronary heart failure undergoing rehabilitation

JIANG Hongmei1,2， CHEN Wenqiang1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 85-88. doi:10.6040/j.issn.1671-7554.0.2013.586

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Objective To investigate the psychological status of elderly patients with coronary heart failure who underwent rehabilitation therapy and psychological intervention. Methods A total of 82 hospitalized elderly patients with coronary heart disease were randomly divided into two groups: the intervention group and control group. All patients received drug therapy for heart failure and rehabilitation exercise training under the guidance of rehabilitation therapist. The patients in the intervention group also received psychological treatments. HAMD and HAMA scores were evaluated. Blood pressure, heart rate, blood glucose and lipid profiles, left ventricular ejection factor (LVEF) and 6-minute walking distance were monitored at the baseline, 4 weeks and 6 months after treatments, respectively. Results There were 53 depression cases (64.6%) and 36 anxiety cases (43.9%) in the two groups. After 4 weeks and 6 months of treatments, the HAMD scores of the intervention group were significantly lower than those of the control group (all P<0.05). Six-minute walking distance of the intervention group was significantly longer than that of the control group after 4 weeks of treatment. Blood pressure, heart rate, lipid profiles, 6-minute walking distance and cardiac function were improved markedly in the intervention group than in the control group after 6 months of treatment. Conclusion Psychological therapy can effectively correct the negative emotion and improve heart function of elderly patients with coronary heart failure undergoing rehabilitation therapy.

Expression and significance of Survivin in human oral squamous cell carcinoma

WU Jinhua1, YANG Xihu1, LIU Zhenzhong2, DING Xiaojun1, XU Jianhui1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 89-92. doi:10.6040/j.issn.1671-7554.0.2013.394

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Objective To investigate the expression of survivin in human oral squamous cell carcinoma and its clinical significance. Methods The survivin expression in 46 cases of human oral squamous cell carcinoma was detected with SP immunohistochemical assay. Results A total of 28 cases (60.8%) presented positive survivin expression, which was closely correlated with metastasis, clinical stages and prognosis. Conclusion Survivin may play an important role in the tumorigenesis and development of human oral squamous cell carcinoma, serving as a valuable factor for its prognosis.

Expression of calpain-2 in human left atrium in rheumatic atrial fibrillation

HAN Bo1,2, WANG Jianchun2, ZHANG Tao2, ZHU Xiaolong2, LI Cong2, WANG Zhengjun2, ZHAO Yong2, ZOU Chengwei2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 93-96. doi:10.6040/j.issn.1671-7554.0.2013.676

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Objective To observe the expression of calpain-2 in the left atrum of patients with rheumatic atrial fibrillation, and to explore its roles in the pathogenesis of atrial fibrillation. Methods A total of 39 patients with rheumatic heart disease who received surgical valve replacement were divided into two groups: the atrial fibrillation group (n=23) and sinus rhythm group (n=16). The expression of calpain-2 was detected with LSAB immunohistochemical and the Real-time PCR technology. Results The deltaCt of calpain-2 mRNA of the atrial fibrillation group (0.091±0.035) was higher than that of the sinus rhythm group (0.066±0.022, P=0.017). Brown particle deposits were observed in the left atrial myocytes in both groups, but the epicardial and endocardial cells showed no brown particles. The average optical density values of calpain-2 of the atrial fibrillation group and sinus rhythm group were (0.92±0.17) and (0.65 ± 0.01)(P<0.001). Calpain-2 levels and left atrial systolic diameter were correlated (r=0.929, P<0.01). Conclusion Calpain-2 exists in the left atrial myocytes. The expression of calpain-2 increases significantly in case of atrial fibrillation.

Effects of PARP-1 on angiogenesis of ovarian cancer

LI Yan, WEI Wei, LV Shuqing, TIAN Yongjie

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 97-101. doi:10.6040/j.issn.1671-7554.0.2013.721

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Objective To investigate the expression of Poly (ADP-ribose) polymerase-1(PARP-1) and its relationship with vascular endothelial growth factor-A (VEGF-A) and microvessel density (MVD) in epithelial ovarian cancer tissues, and to further explore the effect of PARP-1 on the angiogenesis of ovarian cancer. Methods Expressions of PARP-1, VEGF-A and MVD in 60 cases of epithelial ovarian cancer tissues and expression of PARP-1 in 30 cases of normal ovarian tissues were detected by immunohistochemical technology (SP). Ovarian cancer tissues were divided into PARP-1 positive group and PARP-1 negative group with reference to the assessment criteria of immunohistochemical staining. SKOV3 cells were transfected with either negative control siRNA (NC-siRNA group) or PARP-1 siRNA (PARP1-siRNA group), and tube formation assay in vitro was used to determine the angiogenesis capacity. Expression of VEGF-A in supernatants of SKOV3 cells post transfection was tested by ELISA. Results Positive rate of PARP-1 in ovarian cancer tissues (44/60, 73.3%) was significantly higher than that in the normal ovarian tissues (8/30, 26.7%) (P<0.05). In ovarian cancer, expression of PARP-1 was associated with tumor size, histological grade and lymphatic metastasis. Expression of PARP-1 was positively correlated with VEGF-A (P<0.05). MVD value was significantly higher in PARP-1 positive group than in the negative group (P<0.01). Tube density in NC-siRNA group (14.67±1.21) was higher than that in PARP1-siRNA group (8.83±1.47) (P<0.01). Expression of VEGF-A in NC-siRNA group and PARP1-siRNA group was significantly different (P<0.01). Conclusion PARP-1 may promote the angiogenesis of ovarian cancer by up-regulating the expression of VEGF-A.

Potential of breast MRI in predicting the presence of early invasion in ductal carcinoma in situ

HAO Wen, ZHAO Bin, WANG Cuiyan, LI Huihua, QIU Xiuling

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 102-106. doi:10.6040/j.issn.1671-7554.0.2013.671

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Objective To evaluate the potential of breast MRI in predicting the presence of invasion by comparing the morphologic and kinetic characteristics between pure ductal carcinoma in situ (DCIS) and DCIS with early invasion. Methods Preoperative breast MRI of 82 pure DCIS lesions(N1 group)and 51 DCIS lesions with early invasion (N2 group) were reviewed retrospectively. The morphologic characteristics and time-intensity curve (TIC) of all lesions were described in accordance with American College of Radiology Breast Imaging Repot and Data System (ACR BI-RADS). All cases were confirmed by surgery and pathology. Statistical analyses were performed through univariate and multivariate logistic regression approach. Results Lesion size and delayed TIC were associated with the presence of early invasion. Compared with lesion size ＜1.9cm, the probability of invasion was higher for lesion size ≥1.9cm (OR=2.285, 95% CI:1.110-4.663). Compared with persistent TIC, the probability of invasion of washout TIC was 4.773 times (95%CI:1.880-12.118)and the probability of plateau TIC was 2.657 times (95%CI:1.089-6.482). The majority of both pure DCIS and DCIS with early invasion were depicted as non-mass-like enhancement, and there was no significant difference in morphology between two types of lesions(P＞0.05). Conclusion Lesion size and delayed TIC on breast MRI have the potential in predicting the presence of early invasion in DCIS.

Prevalence and characteristics of human papillomavirus infection in pregnant women

LIU Ping1, XU Li2, SUN Yuan1, WANG Zhiping1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2014, 52(4): 107-112. doi:10.6040/j.issn.1671-7554.0.2013.633

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Objective To explore the prevalence and distribution of human papillomavirus (HPV) infection in cervical biopsy tissues or exfoliated cell specimens of pregnant women worldwide. Methods Articles published in PubMed, Medline, Elsevier Science, Web of Science Database and China National Knowledge Infrastructure, Database of Chinese Scientific and Technical Periodicals, Wan Fang Database, and the China Biology Medical Literature Database were retrieved. The overall HPV prevalence in pregnant women, the HPV prevalence in three trimesters and the prevalence of 7 subtypes (16, 6, 18, 11, 58, 31, and 33) were calculated. Results A total of 28 articles were selected, including 13640 pregnant women. Among them, 2294 were HPV positive, with the prevalence rate being 16.82%, and rate among Chinese women being 15.53%. The prevalence rates during early, mid and late pregnancy were 18.20%, 14.38% and 19.32%, respectively. Women younger than 24 years old (23.94%) had the highest prevalence rate. The most common subtypes were 16(3.86%), 6(2.45%), 18(1.80 %) and 11(1.76%). Conclusion HPV prevalence rate is high among pregnant women, especially in women younger than 24 years old, and the most common subtype of infection is HPV-16.

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