Journal of Shandong University (Health Sciences)

Isolation and amplification of human CD4⁺CD25⁺regulatory T cells in vitro

SUN Lei1，WU Jing-jing2，YI Shou-nan2，CHEN Li1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 1.

Abstract ( 1208 )

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Objective To establish a method for isolating and expanding human CD4⁺CD25⁺ regulatory T(Treg) cells in vitro, and to identify the purity, phenotype, activity and function to suppress xenogeneic immune responses of fresh and expanded Treg cells. Methods Human Treg cells were isolated by magnetic activated cell sorting (MACS) and expanded by CD3/CD28 expand beads, IL-2 and rapamycin. Activities and the purity of the fresh and expanded cells were detected by trypan blue staining and FACS, respectively. The phenotype and Foxp3 gene expression were analyzed by FACS and real-time PCR, respectively. The suppression of fresh or expanded human Treg cells was detected by the mixed lymphocyte reaction(MLR) assay. Cytokine-producing cells in coculture were examined by ELISPOT. Results Human CD4⁺CD25⁺Treg cells were expanded up to 3500-fold the number of fresh Treg cells after 2-3 weeks of culture. The activity of the expanded CD4⁺CD25⁺ Treg cells was >97 %, and the purity of them was >96 %. The expanded Treg cells had the similar phenotype and Foxp3 expression with fresh Treg cells. In the coculture experiment (MLR assay), expanded Treg cells were more potent at suppressing CD4⁺CD25⁺- T cell-mediated anti-porcine xenogeneic responses compared with fresh Treg cells, and the difference was significant（71% vs 51%, P<0.05）. The enhanced suppression of expanded Treg cells was associated with increased secretions of IL-4 and IL-10. Conclusions The method established in our study for isolation and expansion of human CD4⁺CD25⁺Treg cells is effective to obtain sufficient cells, which have high purity and activity, and suppressive function uninfluenced.

Influence of long-term ethanol feeding on glycogen synthase kinase-3β and AMP-activated protein kinase in Wistar rats

CHEN Shao-hua1, 2, YU Hui2, 3, WANG Ru-xia4, FENG Li1，2, Zhang Wei2, ZHAO Jia-jun1, GAO Ling5

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 5-10.

Abstract ( 1159 )

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Objective To investigate influences on glycogen synthase kinase-3β(GSK-3β), AMP-activated protein kinase(AMPK) and glucose transporter type 4(GLUT4) after ethanol feeding for a long time. Methods AMPK activator (AICAR) was injected into rats and influences on hepatic glycogen, GSK-3β and AMPK were observed. 48 Wistar rats were randomly divided into 4 groups: feeding different doses of ethanol (A,B,C, 5,2.5, 0.5g/kg·d) or normal saline (D) for 20 weeks. Then GSK-3β, AMPK (AMPKα, P-AMPKα) and GLUT4 proteins, AMPK and GLUT4 mRNAs, and glycogen in the liver were measured. Results AICAR injection resulted in high expressions of glycogen and phosphorylated AMPKα(P-AMPKα) and lower expression of GSK-3β in the liver of rats. In ethanol-fed rats, GSK-3β was over-expressed in the three ethanol feeding groups, especially in group A ,while the P-AMPKα protein expression was significantly decreased(P<0.01). The decreased mRNA and protein expressions of GLUT4 were found(P<0.05). Conclusion Long-time ethanol feeding affected GSK-3β and AMPK levels, and then decreased the level of hepatic glycogen.

Clinical initiation factor of metabolic syndrome

WANG Min1，WANG Lei2， ZHAO Jia-jun3，GAO Ling3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 11.

Abstract ( 1033 )

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Objective To explore the clinical initiation factor of matebolic syndrome(MS). Methods The subjects were selected from male workers on the job at the authorities in Dezhou who had a high prevalence rate of MS. The MS risk factors, such as increased waist circumference(WC≥90cm) and blood pressure (systolic blood pressure≥130mmHg or diastolic blood pressure ≥85mmHg), elevated levels of blood glucose and triglyceride (fasting glucose≥5.6mmol/L, triglyceride≥1.7mmol/L) and decreased level of high-density lipoprotein cholesterol (HDL-C＜1.03mmol/L), were investigated and analyzed among the subjects. All the subjects were divided into two groups: the obese group(WC≥90cm, n=370) and the non-obese group(WC＜90cm, n=228), and the prevalence of MS and characteristics of MS risk factors were compared between the two groups. 136 MS patients selected from the subjects were randomly divided into the diet-alone group (group A, n = 45), the exercise-alone group (group B, n=45) and the diet + exercise group (group C, n=46) for 3-month intervention. The intervention effects were compared and evaluated according to improvements of MS risk factors, oral glucose tolerance test (OGTT) and HOMA-IR among groups A, B and C. Group C, which had a better intervention effect, was divided into two groups according to the improvement of WC after 3-month intervention: group C1 (WC shortening ≥7cm, n=21) and group C2 (WC shortening＜7cm, n=25). Improvements of MS risk factors, OGTT and HOMA-IR were compared between groups C1 and C2. Results MS prevalence of male workers on the job at the authorities was 41.3% in Dezhou. The prevalence of MS increased with age. Increased WC had a high prevalence (62%) and a positive-correlation with increased blood pressure and elevated triglyceride. Blood pressure, blood glucose and triglyceride levels were significantly higher in the obese group at every age stage than those in the non-obese group(P＜0.05). WC, blood pressure, triglyceride, HOMA-IR and OGTT were improved significantly in group C after intervention(P＜0.05). Subjects in group C1 were younger than in group C2(43±10.6 vs 48±9.7, P＜0.05) and blood pressure, triglyceride, HOMA-IR and OGTT in group C1 were significantly improved than those in group C2(P＜0.05). Conclusion Increased WC is a clinical initiation factor of MS. WC control and diet plus exercise intervention can effectively prevent and treat MS, and early intervention has a better effect.

Blood glucose profiles in diabetic NOD mice treated with insulin glargine and aspart

SHAO Peng， LI Gui-mei， ZHANG Li-juan， HU Yan-yan， LIU Cai-hong

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 15-18.

Abstract ( 1050 )

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Objective To evaluate the capacities of insulin glargine and aspart to normalize the blood glucose profile in diabetic NOD mice. Methods Thirty newly diagnosed diabetic female NOD mice were randomized into A,B,C,D and E groups(in each group, n=6). Group A，B , C and D had free access to food and insulin glargine(IG) were given at 9:00 subcutaneously in corresponding doses: 0.1IU/g·d-1、0.05IU/g·d-1、0.015IU/g·d-1 and PBS 5μL·d-1. Intensive insulin therapy was applied in group E, in which insulin glargine, as a basal replacement, was administrated subcutaneously at 9:00, and insulin aspart(IA) was administrated subcutaneously every four hours( beginning at 03：00 every day ) followd by one-hour feeding Meanwhile, another group, i.e.group F(n=6), with the ages parallel to non-diabetic NOD mice, was included in the study. The blood glucose value at different time points (24 hrs) on the fourth day were recorded and the effects of each treatment were analyzed on the 15th day. Results The average blood glucose was significantly lower in group E than Group B,C and D(P≤0.05, respectively). But there was no significant difference found between Group E and Group A/ F(P>0.05). The fluctuation of blood glucose in Group E was significantly smaller than those in group A, B and C(P<0.05, respectively). The percentage of normal blood glucose during 24 hours in group E was much higher than those in group B, C and D(P<0.05, respectively). Severe hypoglycemia and hyperglycemia were often observed in those single insulin glargine groups. The symptoms of diabetes was under control after 15 days of insulin treatment,however, there were 5 and 3 deaths in group A and B due to hypoglycemia. Conclusions Intensive therapy with preprandial IA plus basal IG is superior than once daily basal IG to achieve a better glycemic control.

The morphologic presentation of cochlear in GJB2 conditional knock out mice under scanning electron microscopy

ZHANG Yan-ping1, TONG Ming-wang1, LI Li-na1, ZHANG Xiao-qiang1, SUN Jian-he2, LIN Xi3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 19-22.

Abstract ( 1095 )

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Objective To explore the mechanism of hereditary hearing loss. Methods Cochlea from cCx26 mice on P8（the 8th postnatal day）, P12 （the 12th postnatal day）and P21（the 21st postnatal day） were examined with scanning microscope (SEM). Results On P8 and P12, the external morphology of inner hair cells (IHC), outer hair cells (OHC) and supporting cells developed normally. But in P21 stage, extensive coalesce or absence of stereo cilia of OHC and loss of OHC in the basal and middle turn of cochlea were seen. And the cuticular plate was shriveled and damaged， however， the IHCs in the cochlea were intact without any obvious change. Conclusion Mutation of GJB2 may cause the absence of stereo cilia of OHC, the shrinkage and the damage of Corti′s cuticular plate in mice. All these morphologic changes of cochlea may occur before P21.

Inhibitory effect of JAK inhibitor AG490 on the STAT3 signaling pathway in laryngeal carcinoma

ZHANG Han-bing1, ZHANG Dao-gong2, XIE Guang1, LUAN Xin-yong1, PAN Xin-liang1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 23-27.

Abstract ( 1301 )

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Objective To investigate the influence of JAK inhibitor AG490 on proliferation and apoptosis of human laryngeal cancer Hep-2 cells, reveal the inhibitory effect of AG490 on the JAK/STAT3 signaling pathway, and further explore the potential mechanism so as to study the role of AG490 in the treatment of laryngeal carcinoma. Methods Hep-2 cells cultured in RPMI1640 medium were treated with JAK inhibitor AG490 at different time. The state of cell proliferation was detected by MTT assay, and the apoptosis and cell cycle of Hep-2 were analyzed by flow cytometry(FCW). Expressions of STAT3 and pSTAT3 proteins were detected by Western blot. Results AG490 inhibited cell proliferation in a dose-and time-dependent manner. The apoptosis cells increased with the increase of action time. AG490 resulted in a significant down-regulation in STAT3 and pSTAT3 expressions in the Hep2 cell line. Conclusion AG490 can significantly inhibit proliferation and induce apoptosis of laryngeal cancer Hep-2 cells through decreasing expressions of STAT3 and p-STAT3.

The effect and mechanism of normobaric hyperoxia on blood-brain barrier impairment following cerebral ischemiareperfusion in rats

LIU Bao-yi1, ZHANG Xiao-ming2, GUO Xiao-sun2, HU Wei-cheng2, YUAN Zhong-rui2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 27-31.

Abstract ( 1394 )

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Objective To investigate the effect of normobaric hyperoxia on blood brain barrier impairment following cerebral ischemiareperfusion in rats, and explore the mechanism．Methods 30 normal male SD rats were randomly divided into 2 groups: ① cerebral ischemia-reperfusion control group ( n=15 )， and ② normobaric hyperoxia treatment group ( n= 5 ). Cerebral ischemia-reperfusion rat models were established by the thread ligation of right middle cerebral artery . At 24 hrs after the reperfusion following 90min ischemia, TTC staining, Evans blue dye extravasation and Gelatin zymography were applied to quantify infarction volume, blood-brain barrier impairment and the activity of MMP-9, respectively. Neurological deficit scores were also evaluated. Results Compared with the control group, normobaric hyperoxia treatment significantly decreased Evans blue dye extravasation, the activity of MMP-9 and infarct volume (P﹤0.01) at 24 hrs after the reperfusion. Correspondingly, neurological functions were improved. Conclusions Nnormobaric hyperoxia treatment during the period of focal cerebral ischemia demonstrates the protective effect on blood-brain barrier impairment in the rat model of cerebral ischemia-reperfusion, and the mechanism may be attributed to the suppression of MMP-9.

Effects of honokiol and Bortezomib on proliferation and apoptosis in myeloma KM3 cells

LI Shan, LI Li-zhen, SONG Qiang, ZHAO Chuan-li, YAN Shu-xin, WANG Lu-qun

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 32-36.

Abstract ( 1052 )

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Objective To investigate the anti-proliferative and apoptosis effects induced by Honokiol (HNK) and bortezomib on KM3 human multiple myeloma cell line in vitro． Methods Samples were divided into control group and experimental group, and the latter was further subdivided into HNK treatment group (group B, C, D, E, F, G at respective concentrations of 4,6,8,10,12,15μg / mL), bortezomib treatment group (group H, I, J at respective concentration of 5,10,20μg / mL), and combined treatment group (group K,L, M with HNK 4μg/mL + bortezomib 5ng/mL, HNK4μg/mL + bortezomib 10ng / mL and HNK4μg/mL + bortezomib 20ng/mL, respectively). MTT colorimetric assay were performed to assess the cell proliferation, and cell apoptosis were determined by PI and Annexin V fluorescent staining on flow cytometry. Results Honokiol, at concentrations between 4-15ug/mL, showed significantinhibiting effect on KM3 cell proliferation. At 24h, 48h, and 72h, the inhibition rate rose from 20.38%, 27.45% and 44.94% to 80.54%, 89.99% and 90.91%, respectively. There were significant differences among different groups, or at different time points. The inhibition rates of honokiol on KM3 cells at 48h in H, I, J, K, L, M were 3.13%, 36.22%, 53.99%, 52.68%, 69.99%, 78.53%, respectively. There were considerable differences between K and H, L and I, or M and J (P<0. 05). Apoptosis rate of KM3 cells in B, C, E at 24h and 48h were 6.92%、16.15%、60.70% and 18.84%、37.21%、86.07%, respectively. There were significant differences in apoptosis among different groups at the same time point(P<0. 05). Apoptosis rate of KM3 cells in H and I group were 9.27% and 17.87% at 24h, 11.92% and 53.51% at 48h. Statistical differences were found in apoptosis between H and I group (P<0. 05). Apoptosis rates of KM3 cells in K and L group were 15.75% and 22.18% at 24h, 35.96% and 74.70% at 48h. The apoptosis rates in K and L group were significantly higher than H and I group (P<0. 05). Conclusion Honokiol and bortezomib can inhibit the proliferation of KM3 cells and induce cell apoptosis, moreover, the combination of honokiol and bortezomib can increase the apoptosis rates and enhance cytotoxic activity of KM3 cells.

Indobufen on platelet aggregation and thrombosis

ZHANG Cheng-mei1, YIN Zhi-sheng1, WU Chuan-tao1, JI Jian-bo2, YU Xue-hui1, ZHANG Cheng-e3, LIU Zhao-ping1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 37-45.

Abstract ( 1432 )

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Objective To investigate the effects of Indobufen on platelet aggregation and thrombosis in animal models. Methods In each study, 50 animals were randomly divided into five groups, i.e. group NS, group L(low concentration), group M(medium concentration)， group H (high concentration) of Indobufen and group ASA (Aspirin, positive control）, according to their body weight. After being treated intravenously for 30 minutes, one-minute and five-minute platelets′aggregation rate and the maximum aggregation rate, which were induced by adenosine diphosphate （ADP）, arachidonic acid（AA）or platelet activating factor（PAF）, were assayed. Moreover, the time of thrombotic paralysis or even death caused by collagen-epinephrine and the time of carotid artery thrombosis caused by electrical stimulation during the whole process were tested, too. Results The platelet aggregation study showed that Indobufen group of M, H and ASA 22group, compared with group NS, demonstrated the ability to inhibit ADP, AA and PAF-induced platelet aggregation at different time points（P<0.05 or P<0.01）. In the inducement study of ADP and AA, compared with group ASA, Indobufen group of H showed a significant discrepancy in marked inhabitation of the one-minute/five-minute platelet aggregation rate（P<0.05）. Based on the pulmonary embolism study induced by collagen adrenaline in mice, the survival time of the mice in the Indobufen groups of M， H and group ASA increased much more than group NS（P<0.01 or P<0.05）. Meanwhile, the survival time of group H was considerably longer than group ASA（P<0.05）. According to the results of the carotid artery thrombosis study, Indobufen groups of M，H and group ASA could significantly inhibit the thrombosis time（P<0.05 or P<0.01）， compared with group NS. Although the thrombosis time inhibited by Indobufen did not show a significant difference, Indobufen group of H presented a longer time to inhibit thrombosis than group ASA. Conclusion Indobufen is a very potential inhibitor of platelet aggregation and thrombus formation in vivo, and the effect is dose-dependent. To inhibit the platelet aggregation and thrombosis, Indobufen is better than ASA.

Mechanism of Indirubin to induce the tolerance in immune thrombocytopenic purpura animal model

ZHANG Ai-jun1, HOU Ming2, SUN Nian-zheng1, WEI Wei1, ZHANG Hong1, JU Xiu-li1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 42.

Abstract ( 1057 )

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Objective To study the mechanism of Indirubin on immune thrombocytopenic purpura(ITP) mice model. Method ITP animal model was established with CBA mice received Wastar rat platelet. At the fourth week after the Indirubin treatment, spleen cells isolated from ITP mice was detected with MTT for the proliferation, while the cell levels and phenotypes were observed under FCM. Immunoregulatory function of CD4+CD25+Treg cells isolated by MACS was evaluated with MLR. Result Proliferation of spleen cells in the ITP mice treated with indirubin was significantly inhibited. The ratios of CD4+CD25+Treg cells or CD4+CD25+Foxp3+ Treg cells to CD4+T cells in spleens (P<0.01 ) enhanced by Indirubin were identified. Meanwhile, splenic CD4+CD25+Foxp3+ Treg cells treated with Indirubin showed immunosuppressive ability on the immune response of T effector cells to alloantigens or mitogen as efficiently as the control, CD4+CD25+Treg cells, in vitro. Conclusion The present study indicates that Indirubin can induce the immune tolerance in ITP mice by excerting the effecs on the number and relative proportions of CD4+CD25+Treg cells.

Colon-specific drug release from pH- and time-based pellets of 5-fluorouracil in rats

ZHANG Dong1, LI Cui-jing2, LI Lin-jun1, XI Yan-wei1, HUANG Gui-hua1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 46-50.

Abstract ( 1095 )

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Objective To evaluate colon-specific release properties of pH-and time-based pellets of 5-fluorouracil(5-FU) in vivo. Methods pH-and time-based colon-specific 5-FU pellets were administered to rats by gavage. The rats were sacrificed at different time, and the pellets in different regions of the gastrointestinal tract were collected. Then the morphology of pellets was observed by a scanning electron microscopy, and contents of 5-FU in the pellets and the colonic content were determined by UV and HPLC, respectively. The release characteristics of pellets in different regions of the gastrointestinal tract at different time were evaluated. Results The content of 5-FU was over 95% in pellets collected in the upper gastrointestinal tract before arriving at the cecum. Content of 5-FU could be detected in the colonic content between 5h and 16h, and was up to 84.26% of the total dosage. Conclusion Drug concentration of pH- and time-based 5-FU colon-specific pellets is enhanced in the colon, and the colon-specific effect is exerted by oral administration.

Roles of mitochondrial calcium-activated potassium channel and ATP-sensitive potassium channel in myocardial protection by limb remote preconditioning

CAO Yang1, ZHANG Shi-zhong1, ZHAO Shu-qin2, WANG Xiu-jing1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 51-55.

Abstract ( 1093 )

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Objective To investigate the roles of mitochondrial calcium-activated potassium channel (MitoKCa) and mitochondrial ATP-sensitive potassium channel (MitoKATP) in myocardial protection by limb remote preconditioning (RPC). Methods Seventy-two Male Sprague Dawley rats were divided into nine groups( eight per group): ischemia reperfusion group（I/R）, RPC group, MitoKATP-channel opener group （diazoxide, DZ）, MitoKCa-channel opener group （NS1619）, MitoKCa inhibitor group （RPC+paxilline）, MitoKATP inhibitor group（RPC+5-HD）, RPC+NS1619+5-HD group, RPC+paxilline+DZ group and RPC+paxilline+5-HD group. Preparation of RPC model was made by the ligation of rat femoral artery for 5min followed by 5min reperfusion. The procedures was repeated for 4 times; Preparation of I/R model was made by the ligation of anterior descending coronary artery for 30min followed by 120min reperfusion. Heart hemodynamics of each group was measured. Size of myocardium infarction was measured by TTC staining method and the content of LDH in the effusion from coronary artery was detected spectrophotometrically. Results Compared to I/R group, administration of NS1619(10μmol/L) and DZ(50μmol/L) both improved heart function, decreased myocardium infarction and restrained the release of LDH (P<0.01), which were similar to the remote ischemic preconditioning(RPC) group (P>0.05). However, no difference was observed when compared with RPC group (P>0.05). Administration of paxilline (1μmol/L) or 5HD (100μmol/L) abolished the protective effect of RPC (P<0.01). When one of the two (MitoKCa and MitoKATP) channels was blocked while the other was activated, cardiac protection was still observed. And there was no significant difference when compared to RPC group (P>0.05). When both channels was inhibited by the corresponding inhibitors at the same time, aggravated heart injury was noted compared to the situations in which only one of the two was blocked (P<0.01). Conclusion Opening of both MitoKCa and MitoKATP channels participates in the myocardial protection in limb remote ischemic preconditioning. They may work in independent but cooperative ways.

Mechanism of GABA receptors modulating glutamate receptors

GU Ying-hui, LIU Yong, ZONG Yan-yan, ZHANG Guang-yi, HOU Xiao-yu

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 56-61.

Abstract ( 1292 )

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Objective To investigate the mechanism of γ-aminobutyric acid(GABA) receptors modulating glutamate receptors. Methods Whole-cell patch-clamp recording was used to detect GABA and glutamate receptors-mediated whole-cell currents（IGABA，IGlu） in cultured rat hippocampal neurons. The molecular mechanism of GABA receptors modulating glutamate receptors was studied, using bicuculline（Bic） to inhibit IGABA, and SO2-4 or gluconate- to replace Cl-. Results Co-application of GABA and glutamate-induced IGABA+Glu was less than expected summation of currents IGABA and IGlu, which were induced by application of GABA and glutamate respectively, and the difference was reduced with the increase of concentrations. GABA rapidly and reversibly inhibited IGlu activated by 30μmol/L of glutamate in a concentration-dependent manner. And the inhibition could be reversed by bicuculline, SO2-4 and gluconate-. Conclusion GABAA receptors can down-regulate glutamate receptors, and the mechanism may be related to Cl-permeability.

Construction and characteristics of E-cadherin induced cancerization model from ovarian epithelial cells

ZHANG Hong1, LIU Lian2,3, DONG Ling-ling2,3, DU Chao3, MA Chun-hong3, HAN Li-hui3, LI Li2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 62-66.

Abstract ( 965 )

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Objective To construct E-cadherin induced cancerization model from ovarian epithelial cells and make primary research on its biological behaviors such as proliferation and growth. Methods Chinese hamster ovary (CHO) cells were transfected by plasmid including whole human E-cadherin cDNA and stably screened with G418. Expression of E-cadherin was identified by Western Blot, and cell proliferation test with CCK8 and colony formation test were used to detect their possible malignant transformation of biological behaviors. Results A high level of E-cadherin expression, high ability of proliferation and acquisition of anchorage-independent growth were found in CHO cells transfected by E-cadherin plasmid. Conclusion An E-cadherin induced cancerization model of ovarian epithelial cells was successfully constructed to provide a valuable tool to study the mechanisms of ovarian cancer.

Three dimensional finite element analysis of displacement trends and periodontal stress distribution when a maxillary impacted canine are tracted in different condition

JIN Shu-mei1, WANG Xu-xia1，3, ZHANG Li-na1, REN Xu-sheng1，2, ZHANG Jun1，3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 67-74.

Abstract ( 971 )

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Objective To analyze the the stress of its periodontal tissues in different conditions by establishing the three-dimensional finite element model of an embedded tooth, and provide experimental reference to the orthodontic traction of embedded teeth in the clinic．Methods A patient with a canine completely embedded in the maxillary bone was chosen．The three dimensional finite element model of the canine and its surrounding tissues was established by means of the finite element method (FEM) and spiral CT. On the basis of the dental cusp stable，the canine axis were changed to reach different angles of traction ．Finally，the forces of 50g、100g and 150g were exerted to the three models with different angles at 0°、45° or 90°. Equivalent stress clouds of the periodontal membrane and the wink movement clouds of the tooth were observed. Results At 0°, the wink movement of the embedded tooth was to follow the direction of the force, and the movement was integral. There was a stress concentration area at apical and cusp of the tooth, and the biggest stress emerged at the cusp; At 45° and 90°，the wink movement of the embedded tooth was to follow the traction, but, an angel appeared between the two directions with rotation center at dental cervix. There were stress concentration areas at the apical and the side facing traction forces, and the biggest stress also emerged at the cusp. The stress of periodontal tissues increased following with the growth of the drawing stress at the same traction direction，however, no obvious change of the stress concentration area was found. Conclusions When the traction direction is in line with the tooth axis，the embedded tooth is to move integrally, and the periodontal stress distribution is homogeneous, which benefits the traction. However, when the traction direction angls with the tooth axis, the tooth is inclined to move to one side and the periodontal stress distribution is not homogeneous, which is adversary to the traction. Less drawing stress is suitable to tract the embedded tooth.

Efficacy evaluation of 660nm laser for recurrent oral ulcer

MA Ying-wei1， DONG Li-li1， LIU Tong-jun1， WAN-jun1， HOU Xue-yuan2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 75-77.

Abstract ( 1017 )

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Objective To evaluate the effect of 660nm laser on treating recurrent oral ulcer (ROU). Methods A total of 121 ROU patients (gravistype excluded) were divided into two groups: the experimental group with 61patients and the control group with 60 patients. The experimental group received laser therapy and the control group received drug treatment with Jinda glycerol. The evaluation criteria were DP classification and P-VAS grading. Pain indexes(P) and mean ulcer duration(D) were recorded before treatment, and 3minutes, 5 minutes, 10 minutes and 24 hours after treatment. The results were analyzed by using ttest and χ2 analysis. Results There were significant differences in the pain relief rate between the two groups 3 minutes, 10 minutes and 24 hours after treatment（P＜0.05）, while there was no difference at baseline and 5 minutes after treatment（P>0.05）. There were significant differences in the total effective rate between the two groups （P＜0.05）. Conclusions 660nm laser is more effective in the treatment of ROU compared with the drug, with earlier onset of action and longer duration. It can obviously accelerate healing time and relieve pain.

Microsatellite loss of heterozygosity in primary foci of recurrence laryngeal squamous cell carcinoma and its clinical significance

CUI Zhao-yang1，2， PAN Xin-liang1， WANG Qi-rong2， HAN Fei2， LIANG Hui2， YU Shu-dong2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 78-81.

Abstract ( 1122 )

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Objective To study the characteristic and significance of microsatellite loss of heterozygosity(LOH) in primary foci of recurrence laryngeal squamous cell carcinoma. Methods Allelic deletions analysis was performed on primary carcinoma and adjacent tissues in 31 cases of recurrence laryngeal squamous cell carcinoma with 3 polymorphic markers (D3S1234，D5S592 and D9s171) on 3p, 5p and 9p chromosomal arms by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP)-silver staining. Results Occurrence rates of LOH in the three markers were 48.3%(15/31), 45.2%(14/31) and 61.3%(19/31), respectively. The patients with LOH detected had shorter recurrence time than those with no LOH detected. A statistical significance was found between the two groups(P<0.05). Conclusions There is a negative correlation between LOH and recurrence time. LOH has a predictive value with respect to post-operative relapse in laryngeal cancer patients. It is effectively instructive to choose the best clinical treatment protocol after operation.

Correlation of maspin with angiogenesis and lymphangiogenesis in hypopharyngeal carcinoma

ZOU Ji-dong1, X Wei1, L Zheng-hua1, CAO Hong-yuan1, WANG Hai-bo1, LUAN Xin-yong2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 82.

Abstract ( 1064 )

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Objective To detect the expression of maspin in hypopharyngeal carcinoma and its roles in the development of cancer. And to explore the correlation of maspin with angiogenesis and lymphangiogenesis. Method The expression of maspin protein was examined in 53 patients with hypopharyngeal squamous cell carcinoma by immunohistochemical staining. Microvessel density (MVD) and lymphatic vessel density (LVD) were counted in endothelial cells immunostained with anti-CD105 monoclonal antibody and anti-LYVE-1 multiclone antibody. The correlations of maspin with clinicopathological features, LVD, MVD were analyzed. Result Maspin were found in in 47.2%(25/53) hypopharyngeal carcinoma cases. There were significances among different cell differentiations and between the tumors with or without lymphatic metastasis (P<0.05). The median survival time of positive maspin cases(67 months) was significantly longer then the negative(36 months). In 25 positive cases, the mean MVD was 6.56±2.22 and the mean LVD was 10.76±3.37. As for the 28 negative cases, the mean MVD was 8.14±2.46, and the mean LVD was 15.04±4.57.In the t test analysis, the expression of maspin was negatively correlated with MVD and LVD (P<0.05). Conclusion The reduction of maspin′s expression is correlated with poor differentiation and lymphatic metastasis. In hypopharyngeal carcinoma, maspin may inhibit angiogenesis and lymphangiogenesis.

Detection of coagulation and hemostasis and its significance in patients with recurrent aphthous ulcer

SHANG Hong1, ZHU Yuan-yuan2, WANG Fu-fang1, QI Xiang-min3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 86-89.

Abstract ( 981 )

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Objective To investigate the changes of coagulant and hemostatic parameters in patients with recurrent aphthous ulcer (RAU). Methods 58 patients with RAU and 50 healthy controls were recruited into the study. RAU patients were classified into three groups: the minor aphthous ulcers (MiAU) group, the major aphthous ulcers (MjAU) group and the herpetiform ulcers (HU) group. Factor VⅢ clotting activity (FVⅢ:C), protein C antigen (PC:Ag), total protein S antigen (TPS:Ag), tissue-type plasminogen activator antigen (tPA:Ag), plasminogen activator inhibitor-1 antigen (PAI-1:Ag) and D-dimer were detected by ELISA or by ACL-200 full automatic coagulation analyzer. Results PC:Ag, TPS:Ag, PAI-1:Ag and D-Dimer content were significantly higher in RAU patients than in controls (P＜0.01). The level of t-PA:Ag was significantly decreased in patients with RAU (P＜0.01). PC:Ag was lower in the MjAU group than in the MiAU group (P＜0.05). The lever of FVⅢ:C was significantly increased in MiAU patients compared with the controls (P＜0.01), while no statistical significance was observed between MjAU patients and controls (P＞0.05). Conclusion There were coagulant, anticoagulant and fibrinolytic abnormalities in patients with RAU, and the changes of FVⅢ:C and PC:Ag were correlated with the severity of RAU.

Effect of ultrapure dialysate on erythropoietin resistance in hemodialysis patients

WANG Zun-song1，Zhang Rui-feng2，XU Dong-mei1，CUI Mei-yu1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 90-93.

Abstract ( 1501 )

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Objective To explore the effect of ultrapure dialysate on erythropoietin (EPO) resistance in patients on maintenance hemodialysis. Methods 70 patients taking regular hemodialysis were randomly divided into two groups： the conventional dialysate group（n=35） and the ultrapure dialysate group（n=35）. All the patients had been on hemodialysis for more than 3 months and on the EPO treatment for more than 3 months. EPO dosage was titrated to maintain a target hemoglobin（Hb）level between 110g/L and 120g/L. EPO resistance index (ERI) was calculated as weekly EPO dose/kg weight/g Hb. Laboratory tests including the general examination of blood, biochemistry test of blood, serum ferritin, hypersensitive C reactive protein (hs-CRP) and intact parathyroid hormone (iPTH) were detected in each patient. The relationship between ERI and laboratory factors mentioned above was analyzed. And after 12-month treatment, differences of hs-CRP and ERI were analyzed betweenthe two groups. Results Correlation analysis showed that ERI had a statistically significant correlation with CRP (r=0.432，P<0.01), serum albumin (r =-0.359，P<0.05), iPTH (r=0.316，P<0.05) and nPCR (r =-0.307，P<0.05). hs-CRP was the most important independent predictor of ERI (R2=0.699, P<0.001) by multiple linear stepwise regression analysis. After 12-month treatment, there were marked differences in hs-CRP and ERI between the conventional dialysate group and the ultrapure dialysate group (5.12±2.74 vs 3.77±2.19, P<0.05 and 1.64±0.71 vs 1.10±0.53, P<0.01, respectively). Conclusions CRP, serum albumin, iPTH and nPCR are factors related to ERI . hs-CRP is the most important independent predictor of ERI. Use of ultrapure dialysate should be beneficial to the inflammation status and ERI in long-term hemodialysis patients.

Estimation of dialysis dose (Kt／V Urea) by online clearance monitoring in hemodialysis patients with end stage of renal disease

SUI Zhun, WANG Mei

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 94.

Abstract ( 1524 )

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Objective To investigate the relationship between dialysis dose measured by online clearance monitoring (OCM) and Kt／V Urea from pre-and post-dialysis blood-urea samples, and set up a prediction rule to estimate spKt/V by OCM. Methods OCM and Kt／V Urea data from 74 hemodialysis patients were measured. The relationship between OCM and Kt／V Urea was analyzed by correlation assay. Clinical parameters were put into the linear regression formula, based on which the prediction rule was calculated to estimate Kt／V Urea by OCM. Results Dialysis dose measured by OCM was lower than Kt／V Urea, and they had close relationship（r=0.690，P<0.001）. Based on clinical parameters, Kt／V Urea could be calculated by OCM. Conclusion There is close relationship between OCM and Kt／V Urea. And based on clinical characters, Kt／V Urea could be calculated by OCM.

Combination therapy of transurethral holmium laser resection followed by intravesical instillation of Pirarubicin for non-muscle invasive
bladder cancer: clinical analysis of 52 cases

WU Guang-ping, LI Bo, CAO Yong

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 97-99.

Abstract ( 1189 )

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Objective To summarize the experience of treating non-muscle invasive bladder cancer， using transurethral holmium laser resection of the bladder followed by intravesical instillation of Pirarubicin. Methods From August 2007 to August 2009, 52 patients with non-muscle invasive bladder cancer were treated with the transurethral holmium laser resection of the bladder cancer followed by intravesical chemotherapy of Pirarubicin. Results The tumors were completely resected. The operating time were 15 to 60 minutes, 30 minutes in average. No serious complications, such as bladder perforation during the procedure and bleeding in the perioperative period occurred. All cases were followed for 12 to 24 months after operation, and 5 of the patients ( 9.6%) had recurrent bladder cancer during follow-ups and were retreated with the same method. Conclusion Combination of holmium laser resection of bladder with intravesical chemotherapy of Pirarubicin is an effective and safe therapy, with accurate resection, less bleeding and injuries, earlier recovery and lower recurrence.

State of prethrombosis in the perioperative period of PKRP

WANG Jian-jun, ZHAO Zuo-hui, SUN Bo, JIANG Ting-qi, LI Shun-lai, WANG Wan-li, DONG Gang, WANG Xin-sheng

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 100-103.

Abstract ( 1230 )

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Objective To explore the diagnostic values and relationships of platelet, coagulation, fibrinolytic function and velocity of blood stream in the lower limb deep veins in the state of prethrombosis of benign prostatic hyperplasia (BPH) patients with plasmakinetic resection of the prostate (PKRP) ．Methods 50 BPH patients undergoing PKRP (PKRP group) and 50 non-operative patients (Control group) were enrolled in the study. The plasma platelet (Plt) counting, platelet aggregation index (PAg), platelet adhesion index(PAd), Von Willebrand factor(vWF), prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), fibrogen(Fbg), antithrombin-III (AT-III), protein C (PC), tissuetype plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), and D-Dimer (DD) were measured by automatic blood viscosity dynamic analyser, ELISA or colorimetric methods. Meanwhile, velocity of blood stream in the lower limb deep veins, popliteal vein (Vpop) and femoral vein (Vfem) were measured by color doppler ultrasound．Results In PKRP group, the Pag, Pad, vWF, F1+2, TAT, D-D were significantly higher, while ATⅢ and PC were reduced. Plt, Fbg and PAI-1 were decreased at the beginning but increased later, while the trend of t-PA was just the opposite after the operation. Statistical significances were found(P<0.05)． There was a significant decrease of Vpop and Vfem after the operation compared with those before the operation and the control group (P<0.05). Conclusions Because that Plt aggregation/ adhesion, coagulation system get strengthened, anticoagulation/fibrinolytic system get weakened, and velocity of blood stream in the lower limb deep veins get slowed down,prethrombosis is considrable in the BPH patients with PKRP．

Expressions of Th17, Treg cells and their related cytokines in pre-eclampsia placental tissues

FAN Yue-lian, MA Yu-yan, JIA Xue-qin, TIAN Mei-rong, GUO Yan-yan

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 104-107.

Abstract ( 1137 )

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Objective To investigate expressions of Th17, regulatory T-lymphocytes (Tregs) and their related cytokines in pre-eclampsia and the role in the pathogenesis of preeclampsia. Methods 30 placental tissues of patients with pre-eclampsia and 25 placental tissues of women with normal pregnancy (the control group) were selected. Using realtime fluorescent quantitative PCR, mRNA expressions of the special transcription factor of Tregs Foxp3, Th17-related cytokines IL-17 and retinoic acid receptorrelated orphan receptor C(RORc), regulators for the development and function of Th17 and Tregs IL-23 and IL-27, and IL-27R were quantified, in placental tissues of the pre-eclampsia group and the control group. Results Placenta Foxp3 expression in the pre-eclampsia group was significantly lower than that in the control group（P<0.05）; expressions of Th17-related cytokines RORc and IL-17 in the pre-eclampsia group was significantly higher than those in the control group(P<0.05); also, expressions of IL-23, IL-27 and IL27R in the pre-eclampsia group were obviously increased (P<0.05). Conclusion Th17, Treg cells and their related cytokines are significantly changed in pre-eclampsia, and may involved in the pathogenesis of pre-eclampsia.

Clinical study of treatment discogenic low back pain with CT guided ozone injection

MA Jing-he1，MA Xiao-chun1，WANG Tao1，LANG Ming-lei1， WANG Wei-guo2，XU Dao-zhou3，LONG Hai-bin1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 108-112.

Abstract ( 1228 )

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Objective To explore the effectiveness of the intervertebral disc ozone therapy for discogenic low back pain. Methods From May 2005 to July 2009, 72 patients with discogenic low back pain were treated with CT-guided intervertebral disc ozone injection, of which the concentration and dosage were 30-40 ug/ml and 5-10mL, respectively. 66 cases (91.67%) showed positive pain reproduction. Results All cases were followed up for an average period of 20 months (range from 7 to 54 months). All patients recovered well and returned to their normal lives. Compared with pre-operative period, the average decrease of VAS scoring and ODI scoring were 50.36 ± 2.32（P＜0.01）and 42.23±5.38(P＜0.01）, respectively. Success rate was 90.28%（65/72）and the effective rate was 100%. There were no infection, nerve root injury, or ozone embolism. Conclusion Treatment of discogenic low back pain with CT-guided ozone injection is a safe and effective apporach. CT manifestation of ozone injection is similar to Discography.

High-abundant protein depletion method for cerebrospinal fluid in 2-DE analysis

李芸，杨明冲，刘师莲

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 113.

Abstract ( 1312 )

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Objective To establish and evaluate a method of depleting high-abundant proteins for cerebrospinal fluid (CSF) in two-dimensional electrophoresis(2-DE) analysis. Methods The CSF sample pool was concentrated to an appropriate volume by ultrafiltration, and then high-abundant proteins were depleted with the commercial Albumin and IgG Removal Kit designed for serum. The remaining CSF containing low-abundant proteins was precipitated by acetone and used for 2-DE analysis and 2-DE Western blot identification. Results Most high-abundant proteins could be depleted with this method. The following 2-DE analysis revealed that low-abundant proteins were obviously enriched after the depletion of high-abundant proteins. Compared with the results of 2-DE Western blot, depleting high-abundant proteins enabled the 3 different fragments of tetranectin visible in 2-DE map. Conclusion The high-abundant protein depletion method for CSF established in this study is simply performable, and can be helpful in the detection of low-abundant proteins in 2-DE analysis.

Interventional embolization treatment for intractable epistaxis due to pseudoaneurysm after craniocerebral trauma

GE Ming-xu1, KONG Da-wei1, TENG Hong-tao1, SUN Ping1, PANG Qi2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 117-119.

Abstract ( 1052 )

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Objective To summarize interventional embolization for intractable epistaxis after craniocerebral trauma, and evaluate its effectiveness. Methods Selection of interventional methods for 18 cases was retrospectively analyzed. 10 cases were treated with detachable balloon embolization, 7cases with Guglielmi detachable coil(GDC) embolization, and 1case with trapping because the balloon ruptured repeatedly. Results 8 embolizations were successfully performed with 1 balloon, and the patients′ internal carotid arteries were kept patency. 2 embolizations were performed with 2 balloons, with internal carotid arteries obliterated.6 embolizations were performed with GDC, but the patients′ arteries were kept patency. 1 case of maxillary artery pseudoaneurysm was embolized the artery with GDC. 1 case was treated with trapping of the fistula. After interventional treatment, Digital subtracted angiography (DSA) revealed complete obliteration of fistulas and pseudoaneurysms. In the follow-up of 1 to 5 years, no epistaxis was occurred. Conclusion Interventional embolization is the treatment of choice for traumatic intractable epistaxis. When interventional embolization is failed, trapping of the fistula is an effective therapy.

Effect of APS on TLR4、NF-κB expression in HUVEC-C injured by patients blood with hypertension disease

ZHANG Jing-zhi1，CHENG Li-guo2，HU Xiao-qin3，JIA Hui-xin4

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 120.

Abstract ( 1135 )

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Objective To investigate the effects of APS on TLR4、 NF-κB expression and signal pathway in vascular endothelial cells injured by hypertension patients′ blood. Methods The human umbilical vein endothelial cells (HUVEC-Cs) were intervened by 10% serum of hypertension patient or healthy human for 24h. TLR4mRNA expression in HUVEC-Cs was detected by real time PCR. After the lipopolysaccharide(LPS) activated HUVEC-Cs were effected by different dosage APS, TLR4 and NFκB mRNA were detected by real time PCR, TLR4 and NF-κB protein were detected by western blot. Results After the serum acted the HUVEC-Cs for 24h, TLR4mRNA expression was increased in hypertension disease group than the healthy group(P<0.01). APS showed the ability of reducing TLR4mRNA expression, inhibiting TLR4 protein expression and suppressing IκB protein degeneration in LPS activated HUVEC-Cs in a dose-dependent manner(P<0.05, P<0.01, P<0.001). Conclusion Inflammatory reaction and immunity disorder initiated by TLR-NF-κB signal pathway is one of the influencing mechanisms in the damage of vascular endothelial cells of hypertension disease. APS can reduce the damage.

Effect of Cystatin C on Cathepsin S and extracellular matrix in human arterial smooth muscle cells

WANG Qun1, DU Yi-meng2, DONG Zhao-qiang2, LIU Yu-sheng2, LU Qing-hua2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 124.

Abstract ( 1003 )

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Objective To study the effect of Cystatin C on Cathepsin S and extracellular matrix (ECM) in human arterial smooth muscle cells(ASMCs). Methods ASMCs were divided into the control group(normal cells), the Cystatin C highly expressed group, the Cystatin C silenced group and the independent interference group. Expression of Cathepsin S was detected by RT-PCR and Western blot at different time. Contents of collagenⅠ, laminin(LN) and fibronectin(FN) were detected by ELISA in ASMCs of the control group, the Cystatin C highly expressed group(24h and 48h) and the Cystatin C silenced group(48h and 72h). Results Expression of Cathepsin S in the Cystatin C highly expressed group was lower than in the control group, and the difference between the two groups was not statistically significant（P>0.05）. Expression of Cathepsin S in the Cystatin C silenced group was obviously strengthened with the increase of interference time(P<0.01). While at the same time, expression of Cystatin C was weakened. The result of ELISA showed that content of collagenⅠ in each group was increased or decreased, while the differences were not statistically significant（P>0.05）. Contents ofLN and FN increased in the Cystatin C highly expressed group (24h and 48h), and the differences were both statistically significant compared with the controls(P<0.05). Compared with the control group, content of LN in Cystatin C silenced group (48h) was insignificantly decreased（P>0.05）, while it was obviously decreased in Cystatin C silenced group (72h)（P<0.05）.Compared with the control group, contents of FN in Cystatin C silenced group (48h and 72h) decreased markedly, and the differences were both statistically significant（P<0.05）. Conclusion Expression of Cathepsin S is very low in normal ASMCs. Expressions of Cystatin C and Cathepsin S in ASMCs are inversely correlated. The balance of Cystatin C and Cathepsin S has a significant effect on LN and FN to different degrees, but no significant effect on collagenⅠ.

Diagnostic value of Survivin in pulmonary cancerous pleural effusion

HAO Ke-ke1, ZHU Ji-ping2, YU Li-ke1

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 129-133.

Abstract ( 1069 )

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Objective To investigate the diagnostic value of survivin determination in pulmonary cancerous pleural effusion. Methods The pleural fluid specimens were collected from 102 patients with lung cancer and 87 patients with tuberculous pleurisy. Survivn was determined by ELISA. And the levels of CEA, NSE, and CYFRA211 were detected in the pleural effusion. The results were analyzed with statistical methods. Results The survivn level of the malignant pleural effusion was 52.290±41.421pg/mL, which was significantly higher than that of the tuberculous pleural effusion (17.286±16.734pg/mL)（P<0.01）. Survivin is more sensitive in detemines of pleural effusion than the above other three tumor makers (CEA,NSE, and CYFRA21-1). Conclusion Survivin may be used as a new sensitive and significant marker for diagnosis of lung cancer.

Clinical value of serum CYFRA 21-1, NSE and CEA in the diagnosis and prognosis of lung cancer

SHANG Guan-Hong1，XIAO Wei1, DONG Liang1, LI Yu1, TIAN Hui2, ZHENG Tian-ying3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 134-137.

Abstract ( 3941 )

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Objective To investigate the clinical significance of serum cytokeratin19 fragments (CYFRA 21-1), neuron specific enolase ( NSE) and carcinoembryonic antigen (CEA) in the diagnosis and prognosis of lung cancer． Methods Serum CYFRA21-1, NSE and CEA were measured with chromatometrychemoluminescence method in 68 patients with lung cancer at different TNM stages(the lung cancer group)， 36 patients with benign lung diseases( the lung benign group)， and 28 healthy people(the healthy group), and the relationship of CYFRA21-1, NSE and CEA with TNM stages were analyzed． Survival curves were estimated to analyze the relationship of CYFRA21-1, NSE and CEA with survival time using Kaplan-Meier analysis. Results Serum levels of CYFRA 211, NSE and CEA were significantly higher in the lung cancer group than in lung benign and healthy groups(both P<0.01). There were positive relations of CYFRA 21-1, NSE and CEA with the TNM stage, and negative relations with survival time（P<0.01）. CYFRA21-1 had the highest sensitivity(78.72%) in non-small cell lung cancer, and NSE had the highest sensitivity(91.23%)in small cell lung cancer. Conclusion Serum levels of CYFRA 21-1, NSE and CEA are significantly raised in patients with lung cancer， which suggests that they are important markers for the early diagnosis, development and prognosis of lung cancer．

Influence of personality traits on clinical outcomes in depressive patients

ZHAO Lei1, JIAO Zhi-an2, CHEN Yi-sha3， ZHAO Guo-qing2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 138-142.

Abstract ( 1069 )

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Objective To explore the impact of personality traits of depressive patients on clinical outcomes. Methods Personality traits were evaluated with the Revised NEO Personality Inventory（NEO-PI-R） in depressive outpatients(n=72) and the controls(n=67). The patients were treated with escitalopram for 8 weeks. Severity of depression was assessed before and after treatment by Hamilton Depression Scale (HAMD) and Hamilton Anxiety Scale (HAMA). Results The patients with depression had a significantly different personality profile at the baseline compared with the healthy controls(P＜0.05) and the severity of their illness correlated with high scores in anxiety facet(N1)(r=0.415). The NEO-PI-R showed that N1, N3, C and C3 were strongly associated with score reduced rates of HAMD at the end of 8th week. Conclusion There are significant differences in personality traits between depressive patients and healthy controls. Personality traits may predict the clinical outcome in depressive patients.

Determination of reduced glutathione in the rat livers by HPLC-UVD

CHI De-feng, WANG Xi-feng, SU Guan-min, LI Lin, YANG Yan, SHAO Li-hua

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 142-145.

Abstract ( 1271 )

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Objective To establish a high performance liquid chromatography with UV-absorbance detection method for determination of the level of reduced glutathione in rat livers. Methods After the liver tissue homogenate was made under the condition of ice bathing and precipitated tissue proteins by acetonitrile, the derivative reaction of the supernatant and fluorescent 4-fluoro-7-nitrobenzofuraza was carried out in borate buffer. The mobile phase was acetonitrile-0.02mol/L potassium dihydrogen phosphate (23∶77, V/V), the flow rate was 1.0mL/min, the detection wave-length was 470nm and the injection volume was 10μL. The reaction mixture was quantified with the external standard method. Results The linear range of reduced glutathione was 0.200-125μg/mL (r=0.9999, n=5), and the lowest limit of determination was 0.0400μg/mL(S/N=3). The recovery rate was 96.2%-104.5%, the intra-day and inter-day relative standard deviations were 1.1％-4.6％ and 1.9％-7.5％, respectively. Conclusion This method is simple， rapid, accurate, sensitive and stable, which is suitable for determination of reduced glutathione in the liver.

Preparation and characterization of monoclonal antibodies against deoxynivalenol

ZHAO Li1, WEN Hong-ling1, HOU Xiao-yu2, PU Shuang-shuang1, SONG Yan-yan1, XU Hong-zhi1, Li Feng-qin3

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 146-149.

Abstract ( 1090 )

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Objective To prepare monoclonal antibodies(McAbs) against deoxynivalenol（DON）and characterize their properties. Methods Using a low-dose and longcycle immunization scheme, female BALB/c mice were immunized with DON coupled to bovine serum albumin(DON-BSA). Hybridoma cell lines were obtained by cell fusion. McAbs in the supernatant were determined by ELISA. Hybridoma cells which secreted McAbs to DON-BSA were cloned again and again until the McAb-positive rate reached 100%, and the specificity of McAbs to DON was further confirmed by competitioninhibition ELISA, and a large amount of McAbs was made by ascites in liquid paraffinprimed mice and purified by saturated ammonium sulfate. The subtype of the McAb was tested by a subtype identification ELISA kit. The protein concentration was tested by a BCA kit. The titer, work concentration for reference and affinity constant of the McAb were measured by ELISA. Results The titer of antibodies in sera from Balb/c mice immunized by DON-BSAwas 1∶256000 and all had a strong cross-reaction with BSA. Hybridoma cells which secreted McAbs against DONBSA were screened by ELISA after cell fusion. A hybridoma cell line secreting McAb against DON-BSA was established by 3 rounds of cloning and named 3G5. A large amount of McAbs was obtained by ascites production. The subtype of the McAb belongs to IgG1, the concentration of IgG was 6.06g/L; the McAb titer was 1∶ 500000, the McAb work concentration for reference was 1∶64000 and the McAb affinity constant was 1.62×109. The linear range of the competitive inhibition ELISA calibration curve was from 9.8 to 10000ng/mL, and the linear regression equation was Y=-0.2726X+0.2259(r=0.9309). Conclusion The hybridoma cell line secreting McAbs against deoxynivalenol is obtained, and the prepared antibody showed high sensitivity and specificity to DON, so it can be used to produce a homemade DON-ELISA kit of high quality.

Effects of alum on activities of monoamine oxidase and acetylcholinesterase and content of cholinesterase in the brain of rats

LI Yu-cai, LIU Ping, JIA Xiao-jing, ZHANG Bei-bei, CAO Sheng-nan

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 150-152.

Abstract ( 961 )

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Objective To study effects of alum on monoamine oxidase(MAO) and acetylcholinesterase(AChE) activities, and the acetylcholine(ACh) content in the rat brain. Methods Thirty-two Wistar rats were randomly divided into four groups: the control group, the low dose group, the moderate dose group and the high dose group, and the 3 treatment groups were administrated alum for 30d, the doses of alum in food were 1457, 2914 and 4371mg/kg, respectively. The brain coefficients were calculated. Activities of MAO and AChE, and content of Ach were detected. Results The brain coefficients were increased with the increase of alum in the treatment groups. Compared with the control group, the brain coefficients in the moderate and high dose groups were significantly higher (P<0.01); level of the MAO activity in the high dose group was significantly increased (P<0.05), while the low and moderate dose groups had no significant difference (P>0.05). There was an increasing trend in the AChE activity with increase of alum in the 3 treatment groups, and the AChE activity in the moderate and high dose groups were significantly higher than in the control group (P<0.01). Rats with moderate and high alum administration showed a significant decline in the brain ACh content compared with the control rats (P<0.01). Conclusion Alum can affect activities of MAO and AChE, and content of ACh, and cause damage to the central cholinergic system, which may be one mechanism of alum′s neurotoxic effect on rats.

Preventive effect of garlic oil on cyclophosphamide-induced leukopenia in tumor-bearing mice

LI Yang, YAN Jie, FU Qiang-qiang, ZHAO Sheng, ZHANG Cui-li, YU Li-hua, XIE Ke-qin

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2010, 48(12): 153-157.

Abstract ( 1154 )

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Objective To observe the preventive effect of Garlic oil（GO） on H22 tumorbearing mice in conjuction with cyclophosphamide（CP） chemotherapy. Methods 60 mice were randomized into four groups: tumor model, CP group, 25μg/g GO +CP and 50μg/g GO+CP group. Each mouse was inoculated with Hepatoma 22 cells subcutaneously at the right armpit, followed by 13 days of GO administration. Mice were sacrificed by cervical dislocation, and then tumor mass and spleens were dissected and weighted. In addition, WBC, spleen index, spleen nodes count, the contents of DNA and Nucleated marrow cells (NMC) in bone marrow, bone marrow micronucleus were detected. Results Compared with the tumor model, the tumor mass of CP group decreased by 60.5%, while WBC, spleen index, spleen nodes count, the contents of DNA in bone marrow and NMC were lowered by 65.9%, 34.8%，63.1%，50.1%，64.3%（P<0.05）, respectively. Compared with the CP group, 50μg/g GO caused a tumorinhibiting rate of 62.0%(P>0.05), and increased WBC, spleen index, spleen nodes count, the content of DNA and NMC（P<0.05）. Conclusions Garlic oil can attenuate the leukopenia induced by CP on tumor-bearing mice without influencing its anti-tumor activity.

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