Table of Content

  

16 September 2009
Volume 47 Issue 9

Articles


Articles

Cognition dysfunction of diabetic rats and its relation to apoptosis in the frontal lobe cortex, the hippocampus and HbA1c

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  1-4. 

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To discuss the relationship among cognition function, apoptotic status in the cortex of the frontal lobe and the hippocampus and blood glucose in diabetic rats.  MethodsLearning ability of 26 diabetic rats and 30 normal rats were measured by an electric maze after 12 weeks. Then the rats were executed to obtain the brain tissues to estimate the TUNEL (terminal deoxynucleotidyl transferasemediated dUTP nickendlabeling) positive cells and expressions of Bcl2 and Bax genes in the cortex of the frontal lobe and the hippocampus. Learning ability, the number of apoptotic cells in the frontal cortex and the hippocampus, and glycosylated hemoglobin were analyzed. ResultsLearning ability of diabetic rats was worse than that of the control group, as they needed more time to reach the learning standard(P＜0.001). TUNEL positive cells in the cortex of the frontal lobe and the hippocampus were more in the diabetes group than in the control group. Expression of Bcl2 gene was reduced while that of the Bax gene was increased in diabetic rats (P＜0.01). The time before the diabetic rats reached the learning standard was positively correlated with the apoptotic index of the cortex of the frontal lobe and the hippocampus (r=0.410, P＜0.05), and the learning ability of diabetic rats was positively correlated with HbA1c. ConclusionIncreased apoptosis induced by expression of some regulation genes may contribute to cognition dysfunction in diabetes mellitus.

Clinical characteristics of aging and noaging patients with type 2 diabetes

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  7-10. 

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To compare clinical characteristics of aging and nonaging patients with type 2 diabetes． MethodsClinical information of 285 aging and 282 noaging patients with type 2 diabetes were collected, and disease duration, weight index, blood pressure, fast glucose, serum lipid, glycosylated hemoglobin A1c(GHbA1c) and complications were compared． ResultsThere were significant differences between the two groups in disease  duration, diastolic pressure, and cholesterol (P＜0.05). StandardAchieving rates of clinical index including blood pressure, fast glucose, serum lipid, and GHbA1c in both groups were low, especially in the nonaging group. Incidence of cardiocerebravascular complications, retinopathy and infectious diseases of the aging group were higher than those of the nonaging group (P＜0.05). ConclusionStandardAchieving rates of clinical indexes of the aging and nonaging patients with type 2 diabetes are low, far from the controlling target of treatment guideline, and complications appear ahead of schedule. Prevention and treatment of nonaging diabetes should be enhanced. Cardiocerebrarascular complications, retinopathy and infectious diseases are the focus of treatment for aging diabetes.

Correlation between mild cognitive impairment and advanced glycosylation 
end product level in type 2 diabetic patients

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  11-13. 

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To determine the relationship between advanced glycosylation end products（AGEs) and mild cognitive impairment in type 2 diabetic patients. MethodsBased on the MiniMental State Examination (MMSE) score, type 2 diabetic patients were divided into two groups: one group with 30 cases of cognitive impairment and the other with 32 cases of noncognitive impairment. 30 normal participants served as the control group. AGEs was measured by ELISA in all subjects, also HbA1c, blood lipids, uric acid, fibrinogen, urinary albumin of 24 hours, and other biochemical indicators were measured in all subjects. ResultsThe AGEs level of the cognitive impairment group was significantly higher than that of the noncognitive impairment group (P＜0.05) and the control group(P＜0.01). MMSE score of patients with type 2 diabetes negatively correlated with age， duration of diabetes， HbA1c, body mass index, and serum levels of AGEs（r＝-0.315,-0.276,-0.384,-0.384,-0.282). ConclusionsIncreased AGEs may be an important risk factor for cognitive impairment in type 2 diabetic patients. AGEs may be involved in the pathogenesis of diabetic encephalopathy.

Serum levels of interleukin23, tumor necrosis factorα and interferonγ 
are associated with insulin resistance in type 2 diabetes

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  14-16. 

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To explore the association between serum levels of interleukin23(IL23), tumor necrosis factorα(TNFα) and interferonγ(IFNγ) and insulin resistance in patients with type 2 diabetes. MethodsEightyfive subjects were divided into the normal control group（NGT, 20 patients）, the new type 2 diabetic group (DM1, 33 patients) and the old type 2 diabetic group (DM2, 32 patients). Weight, height, waist circle, hip circle, fasting plasma glucose(FPG), fasting insulin, total cholesterol, triglycerides and HbA1c were determined and serum levels of IL23, TNFα and IFNγ were determined. A simple homeostatic model was used to evaluate insulin resistance(HOMAIR). ResultsSerum IL23 level in groups DM1 and DM2 was increased compared with group NGT(P＜0.05) and was higher in the DM1 group than in the DM2 group（P＜0.01）.　TNFα level in groups DM1 and DM2 was increased compared with NGT(P＜0.05), but there was no significant difference between group DM1 and DM2, also there was no significant difference among the DM1, DM2 and NGT groups in IFNγ level. IL23 level strongly correlated with HOMAIR（r=0.412,　P=0.001）and TNFα（r=0.458，P=0.000）, but did  not correlate with IFNγ（P＞0.05）. TNFα（r=0.410,　P=0.001）and IFNγ(r=0.739, P=0.001）strongly related to HOMAIR. ConclusionSerum IL23 and TNFα levels were significantly increased in type 2 diabetes mellitus and were strongly correlated with HOMAIR. IFNγ level was not significantly different between groups DM and NGT, indicating that IL23 and TNFα were important mediators of insulin resistance in type 2 diabetes mellitus, producing a marked effect without adjusting IFNγ.

CB1 modulates Δ9THCinduced inhibition of LTD in the CA1 area

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  17-20. 

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To explore the role of cannabinoid receptor 1 (CB1) in THCinduced inhibition of LTD in the CA1 area in vitro. MethodsField excitatory postsynaptic potentials (fEPSP) were recorded in the stratum radiatum of the CA1 area in vitro. Results①Low frequency stimulation (1?Hz 15?min) inducedLTD in the CA1 area which was significantly attenuated by Δ9THC; ②THCinduced inhibition of LTD was reversed by SR141716 (a CB1 selective inhibitor); ③ THCinduced inhibition of LTD was not robustly altered in CB1 knockout mice when compared with wildtype littermate mice.  ConclusionCB1 mediates THCinduced inhibition of LTD in the CA1 area in vitro.

Mucociliotoxicity of theophylline/chitosan/βcyclodextrin 
microspheres as a sustained pulmonary delivery

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  21-24. 

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To explore mucociliotoxicity of theophylline/chitosan/βcyclodextrin microspheres as pulmonary sustained drug delivery carriers. MethodsAn acute cell toxicity experiment (LDH and protein in BALF) and a mucociliotoxicity experiment were carried out. ResultsCiliary movement time of MS B microspheres was 512.33 minutes and ciliary beat frequency was 96.0%. The total contents of protein and LDH in BALF of the pulmonary delivery group were not significantly different from those of the control group. ConclusionThese results indicated microspheres possessed better adaptability. 

Antibiotics in reducing resistant mutants of Pseudomonas 
aeruginosa in vitro

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  25-27. 

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To study whether the combinations of two drugs of ciprofloxacinamikacin(CIP), ciprofloxacinceftazidime(AMK) and ceftazidimeamikacin(CAZ) will decrease drugresistant mutants of Pseudomonas aeruginosa in vitro. MethodsMinimum inhibitory concentration (MIC) of CIP,AMK, and CAZ alone was determined by an agar plates dilution method and MIC of the combinations of  them was determined by a checkerboard method. Mutant prevention concentration (MPC) of each single and of combinations were determined by the agar plates dilution method, and then the value of SI(MPC/MIC) was obtained.  Capacity in preventing the appearance of drugresistant mutants was determined based on degree that SI dropped. ResultsSI of CIP, AMk, and CAZ was 16, 16, and ＞32, and of ciprofloxacinamikacin(CIP8; AMK4); ciprofloxacinceftazidime(CIP4; CAZ8); ceftazidimeamikacin(AMK8; CAZ4). ConclusionCompared with each one alone, two drug combinations can decrease MPC and SI to pseudomonas aeruginosa and restrain the drugresistant mutants.

Application of nerve growth factors and fibral glue after 
anastomosis of peripheral nerves in mice

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  28-31. 

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To study the effects of fibrin sealant containing nerve growth factor on peripheral nerve regeneration. Methods40 Kunming mice were randomly divided into two groups: in the processing group the stoma of the nerve was coated by glue membrane mixed with fiberal glue and nerve growth factors （NGFFG）and in the control group only endtoend anastomosis was performed with a 100 suture silk. At 4, 8 and 12?w after the operations, sciatic nerve function index（SFI）, peak of action potential of triceps surae（NAP）and the conduct speed of the sciatic nerve (NCV) were determined. Under microscopy the number of regenerated nerve fibers per view in the stoma was counted and the invasions of fibrous tissues into the stoma were observed. Under  electron microscopy, diameter of regenerated nerve fibers and their axon and thickness of the myelin sheath were measured. ResultsIn each observation stage the sciatic nerve function index,  conduct speed of the sciatic nerve and peak of action potential of triceps surae of the processing group were better than those of  the control group（P＜0.05）. Under microscopy, the number of regenerated nerve fibers per view in the stoma  was more in the processing group than in the control group（P＜0.05）. The incident rate of invasion of fibrous tissues into the stoma in the processing group  was much lower than that in the control group. Under electron microscopy, diameter of regenerated nerve fibers and their axon and thickness of the myelin sheath did not vary between the two groups but were both lower than those of normal nerves, however the maturation degree in the processing group was better than that in the control group. ConclusionsNerve growth factors and fiberal glue used after anastomosis of peripheral nerves prevent invasion of fibrous tissues into the stoma, adhere between the stoma and its surrounding tissues, and effectively step up the growth and regeneration of nerve fibers.

Curative effect and security of Clotridium butyrium on diarrhea mice 
induced by Escherichia coli

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  32-35. 

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To evaluate the curative effect and security of Clotridium butyrium on diarrhea mice caused by Escherichia coli O157;H7 infection.  MethodsDiarrhea  BALB/c mice were respectively treated by Clotridium butyrium, streptomycin, and Bacillus licheniformis. The mortality rates and pathological slides of the small intestine and kidney tissues of mice were observed and compared. Results① The mortality rates of the Clotridium butyrium treatment group were much higher than any other groups. ② The intestinal pathological lesions of mice in the Clotridium butyrium group were lighter than those in the other groups. ③ The pathological lesions in the kidney of mice in the Clotridium butyrium group were much more serious than the other groups. ConclusionClotridium butyrium has some curative effect to improve the intestinal pathological lesions, but aggravates pathological lesions in the kidney, so more attention should be paid to its security.

Changes of TNFα in dental pulp by using distraction 
osteogenesis on orthodontic rapid tooth movement

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  36-39. 

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To observe the changes of TNFα in  dental pulp by using distraction osteogenesis on orthodontic rapid tooth movement. Methods Quantity of TNFαin different stages was detected by ELISA. ResultsThe quantity of TNFαgreatly increased right after four weeks′ distraction force, while it significantly decreased during the first and second week after stopping distraction osteogenesis, and the quantity of TNFαchanged little during day 14 to 28 after stopping distraction osteogenesis, but the TNFαlevel during day 14 to 28 was lower than that of the control group. ConclusionsThe quantity of TNFα could return to the normal level in two weeks after 4 weeks′ distraction force applied to the orthodontic tooth. Day 7 to 14 is the period that the dental pulp  becomes healthy. Orthodontic rapid tooth movement has a slight effect on the dental pulps activity.

Natriuretic peptide in early diagnosis of acute myocardial 
infarction: an experimental study in rats

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  40-43. 

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To explore the changes of brain natriuretic peptide (BNP) concentrations in experimental rats with acute myocardial infarction (AMI) in 14h. MethodsTwentyfive Wistar rats were established for models of AMI and 20 shamoperated rats served as the control group.  The myocardium of the two groups was observed by transmission electron microscopy(TEM) before and after the experiment. Doubleantibody sandwich enzymelinked immunosorbent assay (ELISA) was used to detect serum BNP and cardiac troponin I (cTnI) concentrations 1～4?h before and after the  experiment. ResultsAfter the successful establishment of AMI models 1～4?h, serum BNP concentration was significantly increased compared with that before the experiment and that of the control group (P＜0.01 respectively), and serum BNP concentration at 2h after AMI was the highest(520.66±16.85) ng·L-1. The 14?h serum cTnI concentrations did not significantly vary from the values before AMI and those of the control group  (P＞0.05). Since the establishment of AMI model 14?h, significantly positive correlations were obtained between serum BNP concentrations and the size of AMI, and the highest correlation occurred at 2h after AMI(r=0.69, P＜0.05). No significant differences were noted in serum concentrations of BNP and cTnI in the control group before and after shamoperation (P＞0.05). ConclusionBNP can be used as a blood marker for early diagnosis of AMI especially during 14?h after the onset of AMI and can predict the infarct area.

Effect of electroacupuncture on GAP43 expression in the hippocampus, 
striatum and motor cortex of neonatal asphyxial cerebral palsy rats

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  44-48. 

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To explore the effects of electroacupuncture on the expression of growth associated protein（GAP43）in the hippocampus， striatum and motor cortex  of  neonatal asphyxial cerebral palsy rats, and to explore the significance and mechanism of electroacupuncture in the treatment of cerebral palsy. Methods72 7dSpragueDawley (SD) rats were randomly divided into 3 groups: the sham operation group (n=24), the CP model group (n=24) and the CP+ electroacupuncture group (n=24). The cerebral palsy model was established by occlusion of the left common carotid artery and inhalation of 8％ oxygen plus 92％ nitrogen for 2 hours．　The electroacupuncture group began treatment 24?h after the operations. 6 rats in each group were killed and brains taken on 3, 7,14, and 21?d after the operations. HE staining and immunohistochemical staining (SABC method) were performed. The mean optical density and surface density of GAP43 in the left hippocampus， striatum and motor cortex were determined by ImagePro Plus. ResultsExpression of GAP43 was found at each time point in the sham operation group, which was significantly lower than that in the left hippocampus， striatum, and motor cortex of the model and electroacupuncture groups（P＜0.01）. Expression of GAP43 in the electroacupuncture group was more than that in the model group at most time points with significant differences（P＜0.01 or P＜0.05）. ConclusionElectroacupuncture could promote expression of GAP43 in neonatal asphyxial cerebral palsy rats, accordingly promotes the nerve cell regeneration and synapse reconstitution, which  may contribute to the effect of acupuncture in improving cerebral palsy.

Neruroepithelial stem cells derived from GFP transgenetic mice in vitro

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  49-52. 

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To observe the proliferation and differentiation of neruroepithelial stem cells derived from GFP transgenetic mice. MethodsNeuroepithelial stem cells were isolated from E11d embryos and then amplified by single cell clone and differentiated in serum media. Differentiated cells were identified by immunocytochemistry. Control cells were obtained from nontransgenetic Wistar rats. ResultsThe neruroepithelial stem cells obtained from the GFP transgenetic mice were amplified through single clone and differentiated into neurons and astrocytes in culture media. There was no difference between GFP transgenic cells and control cells. ConclusionGFP transgenetic neruroepithelial stem cells could amplify and differentiate in in vitro culture.

Effect of silencing Notch1 on proliferation of bladder cancer cell line T24

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  53-57. 

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 To investigate the effect of silencing Notch1 gene on cell proliferation in vitro in bladder cancer cell line T24. MethodsThe siRNA eukaryotic expression vector of Notch1（psiRNA1）was constructed using pGenesil plasmid. psiRNA1 was transfected into bladder cancer cell line T24 by lipofectamine. Cell growth, cycle and apoptosis of T24 were determined with methabenzthiazuron (MTT) and flow cytometry (FCM) assay after transfection. The mRNA and protein expressions of Notch1 were determined by RTPCR and western blot. ResultsCompared with the control group, the growth curve of T24 cells was inhibited after transfection, which was timedependent during 60 hours. Notch1 silencing produced a cell cycle arrest at G0/G1（P＜0.05）. The apoptotic cell index also increased to some degree. mRNA and protein expressions of Notch1 were significantly   downregulated
compared with the controls after transfection（P＜0.05）.     Conclusion  Notch1 could inhibit the proliferation of bladder cancer cells by silencing expression of Notch1

, and it might act as a carcinogenic gene in bladder cancer. 

Effect of  glutathione on proliferation of KLE and 
HEC1b endometrial cancer cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  58-61. 

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To investigate the effects of glutathione on proliferation of typeⅠand Ⅱ endometrium cancer cells and to explore its related mechanism. Method① TypeⅠand Ⅱ endometrium cancer cells were challenged by GSH, and proliferation of cells was observed with MTT assay. ② TypeⅠor Ⅱ endometrium cancer cells were treated with 5?mmol/L GSH and the cellular ROS, GSH, and GSSG levels and SOD, CAT and GPx activities were determined. ResultsWithout treatment with GSH, typeⅠ endometrium cancer cells, compared with type Ⅱ endometrium cancer cells, had a lower ROS level and CAT activity and higher GSH level, GSH/GSSG ratio, and SOD and GPx activities. In the condition of treating the cells with reductive GSH, proliferation was promoted in typeⅠendometrium cancer cells while it was inhibited in typeⅡendometrium cancer cells. The ROS level was decreased in both typeⅠand Ⅱ endometrium cancer cells. The GSH level, GSH/GSSG ratio, and SOD, CAT and GPx activities were reduced in type Ⅰ endometrium cancer cells while the GSH and GSSG levels, and CAT and GPx activities were decreased and the GSH/GSSG ratio was markedly increased in type Ⅱ endometrium cancer cells. ConclusionGlutathione definitely differently affects proliferation of typeⅠand Ⅱ endometrium cancer cells, which is related to the ROS level and activities of antioxidative systems in two type of endometrium cancer cells.

Evaluation of androgendependent and androgenindependent 
proteome in LNCaP cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  62-65. 

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To evaluate androgendependent and androgenindependent proteome in LNCaP cells and to investigate the molecular mechanism of androgenindependent growth of prostate cancer. MethodsAndrogensensitive LNCaP parental cells were continuously maintained in a regular cellculture medium. We established a new stable prostate cancer cell (LNCaPs) by culturing androgensensitive LNCaP cells in an androgenreduced condition, and examined the growth properties and androgen responsive of these different LNCaP cells, also observed the morphological changes of those starved cells. We made a comprehensive study of protein expression of the parental androgensensitive prostate cancer LNCaP cells by means of agarose twodimensional gel electrophoresis (2DE) followed by a MALDITOF mass spectrometric analysis. ResultsDuring the creation of  2DE protein maps, 222 proteins in total were successfully identified. Based on strict statistical criteria, 150 spots were found to be significantly differentially expressed in an upand downregulation manner in LNCaPs compared with the parental LNCaP cells, of which we had successfully matched 75 correspondingly. Referring to the LNCaP protein map, it was  found that 26 proteins were upand downregulated in LNCaPs cells, and these proteins were affected by somatostatin treatment. Identified proteins were involved in diverse processes including the stress response, intracellular signaling and apoptosis. Only 7 protein spots in 150 were upregulated in LNCaPs cells. The rest of them were all downregulated. After a qualitative change analysis of protein expression, 6 new proteins were found from 75 presented in the parental LNCaP cells. No new proteins in LNCaPs cell lines  were found compared with parental LNCaP cells. ConclusionIn LNCaP cells, androgendependent and androgenindependent protein differently express and are affected by sms and smsdx. They may provide potential biomarkers for diagnosis, prognosis and therapy targets, while they contribute for prostate cancer protein databases.

Yishen Huoxue decoction regulates FN and COL Ⅳ expressions
through the P38MAPK pathway in mesangial cells 

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  66-69. 

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To investigate the effects of the Yishen Huoxue decoction on secretion of fibronectin(FN) and collgen Ⅳ(COL Ⅳ) through p38 mitogenactivated protein kinase (p38MAPK) in rat mesangial cells. MethodsPharmacologic serum was prepared by the serum pharmacologic method. The cultured mesangial cells in vitro were divided into three groups: the control group(the normal rat blood serum), the stimulate group (the normal rat blood serum+10%IL1β), and the decoction group(traditional Chinese drug +10%IL1β). Levels of FN and COL Ⅳ were determined by ELISA. Changes of phosphated p38MAPK protein andαSMA were determined by Western blot analysis. ResultsCompared with the stimulate group, FN and pp38MAPK expressions of the decoction group significantly decreased（P＜0.01), and expressions ofαSMA and COLⅣ were  also inhibited（P＜0.01). ConclusionYishen Huoxue decoction can decrease expression of αSMA and alleviate levels of FN and COLⅣ, and the p38MAPK pathway may be involved in the mechanism.

Potential relation between atrial fibrillation and angiotensin converting
 enzyme and creactive protein in dog atrial tissues

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  70-72. 

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 To investigate expressions of angiotensin converting enzyme （ACE） and creactive protein （CRP）in dogs with chronic atrial fibrillation（AF）.  MethodsSixteen mongrel dogs were randomly divided into two groups: the pacing group(n=8) and the control group(n=8). High frequency pacing (500±20?ppm) was performed in the pacing group with a permanent pacemaker, while pacing was not performed in the control group. After pacing for 24 weeks, tissue samples were obtained from the left and right atria when dogs were killed. Levels of angiotensin converting enzyme(ACE), creactive protein（CRP） and angiotensin converting enzyme inhibitors(ACEI) in the atria were determined by immunohistochemistry.  ResultsCompared with the control group, expressions of ACE and CRP were significantly enhanced (P<0.01) in atrial tissues during AF, but expression of ACEI was obviously decreased (P<0.01).    ConclusionThere may be relations between the renin angiotensin system(RAS), inflammation and atrial fibrillation, and interruption of the renin angiotensin system (RAS) may inhibit the genesis and development of atrial fibrillation.

Role of the SPK/S1P signal on expression of VEGFmRNA in 
human hepatoma cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  73-75. 

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To investigate the role of the SPK/S1P(sphingosine kinase/sphingosine1phospate)signal on gene level in regulating expression of vascular endothelial growth factor(VEGF) in human hepatoma cells. MethodsThe human hepatoma cell line HepG2 was cultured in vitro. After treatment with a concentration of 520μmol/L DMS(dimethyl sphingosine) in culture media, VEGFmRNA expression was determined by RTPCR. ResultsExpression of VEGFmRNA was obviously inhibited by DMS in a dosedependent manner. ConclusionDMS decreases VEGF expression by inhibiting the SPK/S1P signal, suggesting that the SPK/S1P signal regulates expression of VEGF in hepatoma cells.

Construction of lentiviral recombinants expressing shRNA targeting rPTTG 
gene and evaluation of knocking down efficiency mediated by lentivirus

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  76-80. 

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To construct and screen the shRNAsexpressing lentiviral recombinants targeting the rat PTTG gene. MethodsFour groups of shRNA sequences specifically targeting the rPTTG gene were designed and synthesized. These shRNAs were  inserted into lentiviral vectors pGCLGFP. The recombinants′ transfection efficiency was evaluated after infecting NRK cell line with the lentiviral particles. Real time PCR was employed to assess the gene silencing efficacy of these recombinants. ResultsReal time PCR and sequencing showed that shRNAs were correctly inserted into the lentiviral vector and recombinants were constructed successfully. Transfection efficiency of lenviral recombinants was more than 70%. All of these four shRNAs could achieve gene knockdown effect, and 4# shRNA had the most significant gene silence effect among them(＞80%). ConclusionsshRNAsexpressing lentiviral recombinants targeting the rPTTG gene were successfully constructed and screened. Lentivirus can operated as RNAi vectors to achieve effective transfection. 4# vshRNA could inhibit PTTG expression successfully.  

Construction of coexpression vector pSNAV2.0HSV1tkIREShIL1Ra 
and its expression in synovial cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  81-85. 

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To construct an adenoassociated virus (AAV) vector coexpressing the herpes simplex virus 1 thymidine kinase (HSV1tk) and human interleukin1 receptor antagonist(hIL1Ra) gene by utilizing an internal ribosome entry site(IRES) sequence to link the two cDNAs, and to detect its expression in synovial cells. MethodsThe HSV1tk、 IRES and hIL1Ra were amplified by the polymerase chain reaction(PCR) and inserted into pMD18T simple vector, respectively. After being verified by sequencing, the hIL1Ra and HSV1tk were cloned into pMDIRES to construct pMDHSV1tkIREShIL1Ra. The HSV1tkIREShIL1Ra fragment was then cloned into pSNAV2.0. The recombinant plasmid pSNAV2.0HSV1tkIREShIL1Ra was constructed and identified by restriction enzyme. The recombinant plasmid was transfected into synovial cells with the Lipofectamine 2000 method. The mRNA level change of HSV1tkIREShIL1Ra was identified by reverse transcription polymerase chain reaction(RTPCR). ResultsThe recombinant plasmid was confirmed by restriction endonucleases digestion. The RTPCR results indicated that HSV1tk and hIL1Ra were effectually expressed in the transfected synovial cells. ConclusionThe recombinant plasmid pSNAV2.0HSV1tkIREShIL1Ra was successfully constructed and effectually expressed in the transfected synovial cells, which provided a sound base for gene therapy of osteoarthritis.

Calculating the venous blood concentration of inhalation 
anesthetics with a clinical gas analyzer

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  86-88. 

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ObjectiveTo set up a method to measure blood concentration of inhalation anesthetics using a “headspace equilibration" method with a clinical gas analyzer during surgeries.  MethodsEnflurane, isoflurane, and sevoflurane of six different concentrations were respectively prepared. Voltage peak value of each sample was determined in the mode “GAS SERVICE" of the gas analyzer. Then we got linearity between gas concentration and voltage. The standard blood samples were prepared using the “headspace equilibration" method, and retrieval rate of inhalation anesthetics in blood samples was measured using the clinical gas analyzer and the “headspace equilibration" method. Each sample was repeated six times. Precision of this method, namely relative standard deviations(RSD) was observed.  ResultsThe results were well correlated with the actual concentrations of enflurane, isoflurane, and sevoflurane (r=0.999?8,0.999?9, 0.999?9). The average retrieval rate of enflurane, isoflurane, and sevoflurane was 96.53%, 97.27%, and 97.74%, and their RSD was 1.63%, 1.82%, and 1.24%, respectively. ConclusionA clinical gas  analyzer can be used to exactly measure concentrations of inhalation anesthetics and not increase the error of inhalation anesthetic retrieving rate in the blood.

Stroke risk assessment after transient ischemic attack with the ABCD2 score

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  89-92. 

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To investigate the value of the ABCD2 score in forecasting the risk of stoke after transient ischemic attack (Transient Ischemic Attack, TIA). Methods148 cases of TIA patients were evaluated based on the score of ABCD2 criteria termed by Johnston and the incidence of cerebral infarction was observed within 2, 7 and 90 days after the presence of TIA. ResultsOf the 69 TIA patients whose score was less than 4, 3 had a stroke within 2 days (4.3%), 5 within 7 days (7.2%) and 6 within 90 days (8.7%). Of the 51 patients with a score of 4 or 5, 5 had a stroke within 2 days (9.8 %),8 within 7 days (15.6%) and 10 within 90 days (17.6%). Of the 28 patients at high risk with a score of greater than 5, 8 had a stroke within 2 days (28.6%), 10 within 7 days (35.7%) and 13 within 90 days (46.4 %). The incidence of cerebral infarction of TIA patients with a different score had statistically significant differences（P＜0.05）. ConclusionIncidence of cerebral infarction within 2,7 and 90 days after presence of TIA was related to the ABCD2 score. The higher the score, the higher the incidence. The ABCD2 score is effective and simple for prediction of the short term stroke risk after TIA.

Effects　of TNFα on cultured spinal ligament cells in vitro

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  93-97. 

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To observe expression of tumor necrosis factoralpha (TNFα) in ossification of the posterior longitudinal ligament of the spine (OPLL) and to explore its effect on biological characteristics of ligament cells. MethodsSpinal ligament specimens were obtained from nonossified sites of OPLL patients and nonOPLL patients. Eight OPLL cell lines (OL1OL8) and seven nonOPLL cell lines (NL1NL7) were established. The morphological and osteogenetical characteristics of the ligament cells cultured in vitro were detected.　Expression of TNFαin spinal ligament tissues was determined by an immunohistochemistical (IHC) technique. The effects of rhTNFα in different concentrations on the differentiation of cell lines were also detected. ResultsFour of eight OL cell lines expressed osteoblast phenotypes and the other cell lines expressed fibroblast phenotypes. TNFαexpressed in degenerative and ossified ligament tissues but not in normal or chondrified ligament tissues. TNFαhad different effects on different cell lines, which enhanced the ALP activity, DNA and collagen synthesis of all the cell lines. TNFα also induced OC secretion of all OL cell lines, only two NL cell lines secreted a low level of OC. ConclusionsCultured ligament cells from OPLL patients can express osteoblast phenotypes in vitro. TNFα plays an important role in the formation of OPLL.

Etiologycal and prophylacticotherapeutic analysis of synovial 
chondramatosis around the priximal end of the nail
secondary to internal fixation of  fractures

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  98-100. 

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To research the etiology and the prophylacticotherapeutic measures of synovial chondramatosis around the  end of the nail or wire secondary to internal fixation of  fractures. Methods23 cases of synovial chondramatosis secondary to internal fixation of fractures were analyzed historically,pathologically and clinically.  ResultsSynovial chondramatosis was found in the end of the intromedullary nail in femur shaft fractures in 15 cases，and in the  end of Kirschner wires which fixed the fracture of metacarpus ，metatarsal and  ulna in an other 8 cases.  All the 23 cases of synovial chondramatosis were healed after resection.  ConclusionRepeated mechanical stimulation, overlong detaining period of the nail and the toxic effect of metal may constitute the main causes of synovial chondramatosis. Protective measures are avoidance of overlong detaining period of the nail, an adequate length of backshank and the  use of  latemodel nails.

Studies concerning the effect of fluoride on resistance to erosion of
soft drinks on enamel of primary teeth

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  101-104. 

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To study the effect of fluoride against erosion of soft drinks on enamel surface of primary teeth.  Methods154 pieces of enamel were selected from  retained  human primary teeth and randomly divided into 5groups. After respectively being treated with 0.1%NaF solution, fluoride protector and Bifluorid 12 for 7days, they were immersed at intervals in some beverage for 15 min each time, 5 times per day, for 7days. The SMH and calcium content of all enamel specimens were measured before and after experiments. The data of each enamel slab were analyzed by a repeated measure analysis of variance and Dunn T test. Morphological changes of enamel surface were observed by SEM (scanning electron microscope). ResultsThe SMH and calcium content of enamel slabs obviously decreased after exposure to the beverage. Differences between those treated and nontreated with fluoride was  statistically significant(P<0.001). The Bifluorid 12 and fluoride protector showed a more significant effect than NaF(P<0.05). The loss of gloss, irregular loss of dental tissues and solubilization of enamel primes in variable degrees among experimental groups were found by SEM. After the enamel was treated with fluoride, erosive lesions were less severe. ConclusionSoft drinks have significant erosive effects on enamel of primary teeth. The treatment of enamel with topical fluoride can enhance the resistance of enamel to erosion of soft drinks. This study recommends that Bifluorid 12 and fluoride protector are more efficacious fluorides.

Semiconductor laser in combination with computerIF chronic h ypertrophic pharyngitis in 50 cases

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  105-106. 

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To evaluate the curative effect of MDC semiconductor laser apparatus in combination with computer IF for chronic hypertrophic pharyngitis.  Method150 cases of chronic hypertrophic pharyngitis were randomly divided into the general treatment group, the laser group and the control group.  Results  Symptom scores before and after treatment were statistically different in the general treatment group(P<0.01), and the effect rose on week two and peaked on weeks 4 to 8. However the symptom scores were not significantly different between the laser and control groups (P>0.05) and were significantly higher in the general treatment group than in the laser and control groups. ConclusionMDC semiconductor laser therapy in combination with computer IF achieves a curative effect on chronic hypertrophic pharyngitis.

Expression of regulated upon activation normal T cells and secreted 
factor(RANTES) and its receptor (CCR3) in oral lichen planus

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  107-110. 

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To detect expressions of RANTES, CCR3 and RANTES mRNA in the tissues of oral lichen planus (OLP) and normal oral mucosa and to explore the role of RANTES and CCR3 in the process of OLP. MethodsSpecimens were collected from 30 patients with OLP and 10 patients with normal oral mucosa. Expressions of RANTES and CCR3 were determined by the streptavidinperoxidase（SP）technique. Expression of RANTES at the level of mRNA was observed by ISH. ResultsExpressions of RANTES and CCR3 in OLP were significantly higher than those in normal oral mucosa. ConclusionCompared with tissues of normal oral mucosa, different expressions and distribution of RANTES and CCR3 in OLP suggest that RANTES and CCR3 probably play important roles in the process of OLP.

CT cisternography in the classification and treatment of 
childhood intracranial arachnoid cyst　

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  111-113. 

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To assess the application value of CT cisternography (CTC) in the diagnosis and treatment of childhood intracranial arachnoid cyst (IAC). MethodsCTC was performed on 19 patients with intracranial arachnoid cysts to identify whether the cysts were communicating. NCIAC accompanied with apparent clinical symptoms or dysplasia of local brain lobe served as indications of operations. After the operations, patients were given CTC again to evaluate the therapeutic effect. Results6 cases had communicating intracranial arachnoid cysts (CIAC), and 13 cases had noncommunicating intracranial arachnoid cysts (NCIAC). Postoperative CTC demonstrated that all the cysts communicated  well with the cistern. ConclusionCTC is a simple and reliable method for classification and diagnosis of childhood IAC, but it is a traumatic examination for patients.

Relationship between single nucleotide polymorphism of TRAIL gene
 promoter region and fatty liver disease

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  114-118. 

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To investigate the distribution of single nucleotide polymorphism ( SNP) in the promoter region of tumor necrosis factorrelated apoptosis inducing ligand (TRAIL)gene and to evaluate the effect of  SNP in the promoter region of the TRAIL gene and FLD susceptibility in the Chinese Han population from Shandong Province. MethodsGenotype and allele frequencies of SNPs -707C/T, -665T/C, -621C/T and -597A/G in 42 and 67 nonor alcoholic fatty liver disease and 132 healthy controls were detected by polymerase chain reaction and restriction fragment length polymorphism (PCRRFLP) and Tetraprimer ARMSPCR methods, and then analyzed with Chisquare tests. ResultsNo SNPs at -707, -665, -597 sites were found in the Chinese Han population from Shandong Province. Frequencies of CC/CT/TT genotypes at -621 site were 0.238/0.500/0.262, 0.269/0.448/0.284 and 0.280/0.394/0.326 separately in the non or alcoholic fatty liver disease patients and healthy controls. However, genotype and allele frequencies of this site showed no significant difference among different groups. ConclusionThere were no SNPs at -707,-665 and -597 sites. SNP of -621 site in the TRAIL gene promoter region showed no association with FLD susceptibility in the Chinese Han population from Shandong Province.

Colonoscopy training with a computer～based simulator

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  119-121. 

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o explore the role of a computerbased simulator in colonoscopy training. MethodsThirtyseven persons with no experience in colonoscopy were randomly assigned to two groups: one group underwent 10 hours of training with a computerbased simulator, and the other did not. Each trainee performed colonoscopy in 20 patients. Performance parameters evaluated cecal intubation and procedure duration. ResultsDifferences were significant for procedure duration（8.51±2.82 vs 10.15±3.22，P<0.05）and  cecal intubation（214/380 vs 141/360，P<0.05）. ConclusionComputerbased simulators are effective in providing trainees with skills needed for colonoscopy.

Effect of miniinvasive hematoma aspiration on the content of glial

fibrillary acidic protein in patients with cerebral hemorrhage

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  122-124. 

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To study the concentration of serum glial fibrillary acidic protein（GFAP）in patients with hypertensive cerebral hemorrhage after minimally invasive surgery (MIS). MethodsBlood samples were obtained on day 1 (before the operation in the MIS  group), 3, 7, and 14 after the onset in 74 patients with MIS and 80 patients with non minimally invasive surgery (NMIS), and 60 normal controls(the NC group). Serum GFAP was detected by enzyme linked immunosorbent assay(ELISA). ResultsThe content of serum GFAP in the NMIS group significantly increased on day1, rose to a climax on day7, and then decreased, but was still significantly higher than that in the control group. Compared with the NMIS group, the level of serum GFAP in the MIS group significantly decreased on day 7 and 14. GFAP  had correlations with nerve function deficiency score (NDS). ConclusionsMiniinvasive hematoma aspiration can decrease the serum level of  GFAP in patients with cerebral hemorrhage. Serum GFAP is a potential marker for prognosis and outcome in patients with nonminimally invasive surgery.

Epidemiological and clinical characteristics of mycoplasm pneumonia (MP) 
infection in communityacquired lower respiratory tract
infection among hospitalized children

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  125-128. 

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To summarize the epidemiological and clinical characteristics of mycoplasm pneumonia (MP) infection in communityacquired lower respiratory tract infection (CALRTI) among hospitalized children. MethodsA retrospective analysis of a database of 876 patients with CALRTI at the Qilu Hospital of Shandong University between 2007 and 2008 was performed. Serologic diagnosis for MP infection was made based on single titers ≥1∶80, which was measured by a passive particle agglutination test. ResultsMP infection was diagnosed in 177 patients aged from 6 months to 14 years. The mean age was 5.1 years. Children older than 3 years accounted for 79.7％. The frequency of titer of MPIgM ≥1:320 was significantly higher in patients older than 3 years than in those younger than 3 (88/119 vs 25/58, 73.9% vs 43.1%). Prolonged cough and fever were leading symptoms. In children younger than 3 years, moderate fever and wheezes on auscultation are the main clinical features. Chest Xray showed typical radiological features such as peribronchial and perivascular interstitial infiltrates, with very few massive infiltrations. Extrarespiratory complications were rare.  In children older than 3 years, high fever and persistent dry cough were prominent features with less manifestation on auscultation. Massive infiltrations  and pleural infusion were often seen on chest Xray and extrarespiratory complications were common. ConclusionsMycoplasma pneumonia infection is frequent in children over 3 years among hospitalized CALRTI patients. The infective rate increases as the age increases. The infective rate in summer is significantly lower than that in spring, autumn and winter. The clinical features are obviously different between patients older and younger than 3 years.

Characteristics of microvessels and mature vessels in hepatocellular 
carcinoma and their relationship

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  129-133. 

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 To explore differences in characteristics of microvessels and mature vessels of hepatocellular carcinoma (HCC) and to investigate their relationship. MethodsThirtyone cases of HCC proved by pathology were enrolled in this study. Continuous slices were stained by immunohistochemistry for determination of vascular endothelial growth factor (VEGF) and its receptor (Flk1), proliferating cell nuclear antigen (PCNA), CD34 and αsmooth muscle actin (SMA). Expressions of VEGF， Flk1, and PCNA were determined and vascular parameters such as number, mean area, overall area, circumference, diameter, distance among adjacent vessels (DAV), and variety indexes of microvessels and mature vessels were counted. Arterioles and veinlets, mature vessel indexes, mean perfused fraction (mPF), extracellular space (ES) and extracellular extravascular space (EES) were also calculated.  ResultsNumber, overall area and variety indexes of microvessels were more than those of mature vessels, whereas mean area, diameter and DAV were less than those of mature vessels (P＜0.05). Number, circumference, and ES were different in microvessels between positive and negative VEGF expressive groups, whereas they were not different in mature vessels (P＞0.05). Number, circumference and variety indexes of microvessels and arterioles were different among different Flk1 groups (P＜0.05). In addition, PCNA was positively correlated with variety indexes of microvessels (P=0.045, r=0.563), and negatively with DAV ConclusionThere vascular characteristics between microvessels and mature vessels. VEGF and its receptor, Flk1, may regulate the angiogenesis of microvessels, but they have few effects on mature vessels.

Effect of systematic prenatal health education and appropriate 
prenatal diagnosis on pregnancy outcome

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  134-136. 

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 To analyze the effect of systematic prenatal health education and an appropriate prenatal diagnosis on pregnancy outcome. Methods266 women who had undergone prenatal health education and prenatal diagnosis were enrolled in the experimental group, and 208 pregnant women who had not undergone these served as the control group. Their knowledge about antenatal care, delivery, infant care and their pregnancy outcome were analyzed. ResultsThere was significant deviation(P＜0.05) in their knowledge about pregnancy nutrition, cesarean advantages and disadvantages, normal delivery, and infant care. The control group had a higher cesarean section rate than the experimental group(P＜0.05). ConclusionSystematic prenatal health education and an appropriate prenatal diagnosis for pregnant women obviously improve maternal health and pregnancy outcome, and decrease the mother and fetus disease rate.

Prevalence of prehypertension and its influential factors  among railway workers

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(9):  137-140. 

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To analyze the prevalent rate of prehypertension and its influential factors for railway workers. MethodsCluster sampling was used to select 12?094 health physical examination results among railway workers. Single factor analysis and multivariate logistic analysis were used. ResultsThe prevalent rate of prehypertension among railway was 47.2%, which was significantly different in different age and gender groups. Single factor analysis showed that smoking, alcohol, FPG, high TG, and high BMI were risk factors of prehypertension. Multivariateregression analysis showed that gender, age, smoking, alcohol, FPG abnormality, high TG and high BMI were risk factors of prehypertension. ConclusionThe prevalent rate of prehypertension is at a high level and prevention and treatment of hypertension among railway workers should be strengthened.