Table of Content

  

16 November 2009
Volume 47 Issue 11

Articles


Articles

Effect of MPO on monocyte chemoattractant protein1 in endothelial cells and the intervention of metformin

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  1-5. 

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To observe the effect of myeloperoxidase (MPO) on monocyte chemoattractant protein 1(MCP1) expression in endothelial cells and the intervention of metformin. MethodsMCP1 and AMPKα protein expressions in endothelial cells were determined by the westernblot method, and the level of MCP1 in  culture medium was measured by the sandwich ELISA method. Also, RNA interference technology was used to determine the role of AMPKα. ResultsBoth MCP1 protein expression in endothelial cells and content of MCP1 in culture medium were significantly increased by MPO in a dose and timedependent manner（P＜0.01）, and metformin decreased  MCP1 protein expression and content of MCP1 by promoting phosphorylation of AMPKα, however, AMPKαSiRNA significantly weakened the effect of metformin（P＜0.01）. ConclusionMetformin can inhibit the effect of MPO on endothelial cell MCP1 protein expression and MCP1 concentration.

Expression of visfatin mRNA in adipose tissues of subjects with different glucose tolerance

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  6-8. 

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To investigate the difference of visfatin mRNA expression  in abdominal subcutaneous and omental adipose tissues of  type 2 diabetes and impaired glucose tolerance(the IGT group) and to explore the relationship between visfatin mRNA expression and indexes of correlation metabolism. MethodsThe subjects were Chinese Han population, including 23 controls with no family history of diabetes(the control group), 18 patients with impaired glucose tolerance(the IGT group) and 22 with type 2 diabetes(T2DM group).  The blood pressure, FBG, TC, TG, HDLC, LDLC, HbA1c, and Fins were determined, and (T2DM BMI, WHR, and HOMAIR were calculated. Semiquantitative RTPCR was used to determine  visfatin mRNA expression in adipose tissues. ResultsVisfatin mRNA expression of omental adipose tissues in the T2DM group was higher than that in the IGT and control groups(P＜0.01), but was not significantly different between the IGI group and the control group (P＞0.05). There was no statistical difference in visfatin mRNA expression of subcutaneous adipose tissues among the three groups (P＞0.05). Visfatin expression of omental adipose tissues in the fat/overweight group was higher than that in the control group (P＜0.01). Visfatin mRNA expression of omental adipose tissues was higher than that of subcutaneous adipose tissues (P＜0.01). Correlated analysis showed that visfatin expression of omental a adipose  tissues had a relation to WHR(P＜0.05), but not to  age, BMI,　SBP,　DBP,　FINS,　FBG,　HbA1C, and HOMAIR. While visfatin mRNA expression of subcutaneous adipose tissues was related to TC(P＜0.05). ConclusionVisfatin expression increases in both T2DM patients and fat people, mainly in the omental adipose tissues, and plays effects in maintaining the balance of glucose homeostasis.

Platelet function in subjects with impaired glucose tolerance

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

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To investigate the state of platelet activation and its influencing factors in subjects with impaired glucose tolerance(IGT). Methods33 patients with IGT were enrolled in this study. 30 DM and healthy persons served as the controls. Percentage and mean fluorescence intensity of CD62P, PAC1 as indexes of platelet activation were measured using whole blood flow cytometry (FCM). Changes of count of blood platelet, mean platelet volume(MPV), platelet distribution width (PDW) and plateletlarge cell ratio(PLCR)were analyzed with a blood cell counter. TC, TG, HDLC, LDLC, VLDLC, FPG and FIns were simultaneously measured. ResultsIGT and subjects with diabetes had higher levels of CD62P%(P＜0.01) and CD62MFI(P＜0.05) and PAC1% (P＜0.01) compared with the NC group. CD62P%,CD62MFI and PAC1% had no statistical differences between the IGT and DM groups(P＞0.05). By stepwise regression analysis, CD62P in the IGT group had relation to PLCR, LDLC, TC and body mass index (BMI) independently. PLCR, MPV and DBP were contributors to PAC1. ConclusionThere is the  same degree of platelet activation in subjects with IGT, and DM. LDLC, TC, DBP and BMI are contributors to CD62P and PAC1 expressions on platelet in IGT.

Clinical research of chronic obstructive pulmonary disease  and insulin resistance

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  14-16. 

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To  investigate the relativity of chronic obstructive pulmonary disease（COPD） and insulin resistance（IR）. MethodsA total of 330 cases of COPD were divided into four groups based on the chronic obstructive pulmonary disease diagnosis and treatment guidelines (revised edition 2007): 95 cases of  mild COPD, 95 cases of moderate COPD, 75 cases of severe COPD, 65 cases of extremely severe COPD. 40 persons served as the normal controls. Fasting plasma glucose and insulin, blood lipids, urine albumin excretion rate, and insulin sensitivity index(ISI) were determined. ResultsCompared with the normal control group, fasting blood glucose, blood lipids, urinary albumin excretion rate in different COPD groups were significantly increased, while ISI was markedly reduced(P＜0.05). Compared with the mild and moderate COPD groups, fasting blood glucose, blood lipids, urinary albumin excretion rate of the severe and very severe COPD groups were significantly increased, while ISI was significantly reduced (P＜0.05). ConclusionPatients with chronic obstructive pulmonary disease have insulin resistance, and the more serious the disease, the lower the insulin sensitivity.

Metabolism ofβamyloid precursor protein, expressions of its related enzymes and effect of pioglitazone intervention in the brain of
insulin resistance rats

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  17-20. 

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To investigate the expressions of βamyloid (Aβ), amyloid precursor protein (APP) and its metabolismrelated enzymes, and to explore the effect of pioglitazone (PIO) intervention in the brain of insulin resistance rats.  MethodsOf 45 Wistar male rats, 10 were randomly chosen as the control group (NC group), and the others were given 10% fructose for 4 weeks to develop the insulin resistance (IR) model. 26 IR rats were randomly divided into the IR group(n=13) and the PIO group (n=13). The PIO group was given pioglitazone(10?mg·kg-1·d-1 by gavage for 12 weeks, and the IR and NC groups were given an identical volume of physiological saline. Immunohistochemistry and Western blotting were employed to examine the level of Aβ42 in the hippocampus and the changes of APP, βsecretase (BACE1) andγsecretase (PS1) in the brain tissue.  ResultsImmunohistochemistry results indicated that the optical density of Aβ42 in the hippocampus of the IR and PIO groups was significantly higher than that of the NC group, and was lower in the PIO group than that of the IR group(P＜0.01). Western blotting showed that APP, BACE1 and PS1 levels in the rat brain were elevated in the IR and PIO groups compared with that in the normal controls，and the concentration in the PIO group was lower than that of the IR group (P＜0.05). ConclusionInsulin resistance promotes Aβ42 by upregulating the activities of BACE1, PS1 and pioglitazone inhibits the activities of BACE1 and PS1 to decrease the expression of Aβ42.

Cloning human βcatenin gene promoter and analyzing its activity

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  21-24. 

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To clone a 1.8?kb fragment upstream of the βcatenin gene and assay its promoter activity． MethodsA 1.8?kb fragment upstream of the βcatenin gene was amplified by PCR using human genomic DNA as a template． Its promoter activity was determined with dualluciferase reporter assay after it had been cloned into a pGL3basic vector and transfected into PC3 cells． ResultsThe sequence of the 1.8 kb fragment proved to be correct by DNA sequencing． Dualluciferase reporter assay (Ml／M2) was 11.71 at 48?h after PGL31.8?kb was cotransfected with pRLTK into prostate cancer cell PC3 which was about 2.43fold higher than that of pGL3control cotransfection with pRLTK,  206.31 fold higher than that of pGL3basic cotransfection with pRLTK and 21.38 fold higher than that of pGL3promoter cotransfection with pRLTK. ConclusionThe cloned 1.8?kb fragment upstream of the βcatenin gene presented  strong promoter activity．

Effect of p38MAPK inhibitors on early development and implantation in mouse embryo

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  25-28. 

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To observe the biological effect of p38 MAPK inhibitors on early   development and implantation in mouse embryos. Methods① Embryo culture in vitro：  early mouse embryos were  randomly divided into groups and cultured with 0.2, 2, and 20?μmol SB203580. ② Intrauterus injection in vivo: SB203580 of different concentrations was injected into the uterus on day 4 of pregnancy. The implantation number was calculated and the histological changes were examined on day 8 of pregnancy. Results① In the inhibitortreated groups, embryos did not develop to the blastocyst and were halted at the 8 to 16cell stage. The treated embryos remained viable as the developmental blockage was rescued by removing embryos from the drug treatment and placing them in a drugfree medium until they progressed to the blastocyst stage. ② The average number of embryos implanted in the p38MAPK inhibitor treated uterus was significantly lower than that in the control uterus (P＜0.01). Microscopic observation demonstrated that the embryo was degenerated and even necrosed in some parts. In the decidual cells there was obvious adipose degeneration, while many blood cells were invaded in the embryos and the deciduas. Conclusion ① p38MAPK activity may be required to support  embryo development through the murine preimplantation interval. ②  p38MAPK inhibitors can inhibit the blastocyst implantation and the development of embryos and decidual cells in vivo.

Effects of Dishevelled2 and Vangl2 on the development of cleft palate induced by excess retinoid acid in KM mice

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  29-33. 

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To explore the effects of Dishevelled2 and Vangl2 protein on cleft palate induced by excess retinoid acid in KM mice. MethodsIn both malformed embryos induced by excess retinoid acid (the experimental group) and normal KM mice (the control group),  expressions of Dishevelled2 and Vangl2 in the epithelium of the palatal shelves and the mesenchyma were determined by  immunohistochemistry and image analysis. ResultsExpression of Dishevelled2 in the mesenchyma of the experimental group was significantly higher than that in the control group, however that in the epithelium of the palatal  shelves of the experimental group was significantly higher, but then lower than that in the control group. So was expression of Vanlg2 in the mesenchyma. ConclusionExcess retinoid acid may interfere with normal expression of Dishevelled2 and Vangle protein in developing palates and causes cleft palate in KM mice.

Effect of Edaravone on cognition ERK1/2 activation and Cytc
release after traumatic brain injury in rats

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  34-37. 

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Abstract： ObjectiveTo study the effects of Edaravone on cognitive  function and its potential mechanism after traumatic brain injury. MethodsMale SpragueDawley rats were randomly divided into the control group（n=40），　the traumatic group（n=68），　and the Edaravone treatment group（68）. TBI rat models were established based on description of the Marmarou′s diffused brain injuries. At 1，6，24，48，72?h after injuries， the histopathological changes of mitochondria were observed with an electron microscope. Expressions of pERK1/2 and Cytc were determined by immunohistochemistry and WesternBlot. Learning and memory functions (Morris water maze) were determined daily 3?d after injuries for 7?d. ResultsAfter TBI， some mitochondria displayed swelling, spinal fracture, or even disappeared. Expression of pERK1/2 increased with the development of TBI and peaked at 24 h, then gradually decreased and expression of Cytc increased and peaked at 48?h. The searching safety island period山of rats in the traumatic group(230.9±20.9) was significantly longer than that of rats in the control group(50.7±4.9). While the histopathological damages of mitochondria and expressions of pERK1/2 and Cytc were significant decreased in the Edaravone treatment group, and the searching safety island period (130.0±30.8) was markedly restored(P＜0.05). ConclusionEdaravone has good therapeutic effect on traumatic brain injury and the molecular mechanism is related to attenuating ERK1/2 activation and Cytc release following the trauma.

Effects of Adrenomedullin on rats with early trauma in the kidney

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  38-41. 

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To investigate the effects of extragenous Adrenomedullin (ADM) on the score of the renal tubule in rats with early renal mechanical trauma. Methods104 healthy adult plain grade Wister rats were randomly divided into four groups:8 in the control group, 32 in the trauma group, 32 in the group treated with ADM before trauma, and 32 in the group treated with ADM after trauma. The experimental models were prepared by free dropping a ferrous hammer in the last three groups. ADM（0.1?nmol/kg） was intrapertoneally administrated 10 minutes before trauma or after trauma in the ADM groups. Renal tissues  determined the pathotransform changes by HE staining. ResultsThe score of the renal tubule was distinctively increased in the early phase (1?h and 6?h) (P＜0.05) but significantly decreased to the level of the control group in the late phase post trauma(12?h and 24?h)(P＞0.05). ConclusionExtragenous ADM can protect the kidney, which has relations to the time of intervention.

Inhibition of Radix Astragali to expressions of αSMA and FN in the lung tissues of mice with asthma

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  42-45. 

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To investigate the inhibition of Radix Astragali′s to expressions of αSMA and FN in the lung tissues of mice with asthma. MethodsThe BALB/C mice were divided into three groups: the control group(Group A, n=20), the asthmatic model group(Group B,　n=20), and the Radix Astragali managed group (Group C, n=20). A mouse model of asthma was established by exposing the animal to aerosolized ovalbumin (OVA). The managed group was given Radix Astragali before every stimulation. After being sensitized, the mice  were subjected to laboratory tests. Expressions of FN mRNA and αSMA mRNA of the lung tissues were determined by RTPCR and AlphaImager2200 semiquantitation analysis system. Masculine expressions of FN and αSMA were determined by twostep immunohistochemistry and leica QWIN V3 analysis system.  Result①Compared with those in group A, expressions of αSMA and FN in group B were significantly increased (P＜0.01). Compared with group B, those in group C were significantly　 decreased (P＜0.01).　②Compared with those in group A, expression levels of αSMA mRNA and FN mRNA in group B had a great increase (P＜0.01).　There was a significant decrease of αSMA mRNA and FN mRNA levels in group C compared with group B(P＜0.01).　ConclusionFN and αSMA are important symbols of airway remodeling. Radix Astragali can inhibit expressions  of αSMA and FN in the lung tissues of mice with chronic asthma, and may inhibit airway remodeling in asthma.

hFGF7 related modeling peptide screening with phage random twelve peptide library

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  46-49. 

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To harvest hFGF7 modeling peptides from a phage random twelve peptide library to promote epidermal cells proliferation. MethodsA phage random twelve peptide library was screened 4 times by monoclonal antihuman FGF7. ELISA staining examination was performed to choose monoclonal phages at random. Also, DNA extracted from phages with better bindingactivity was sequenced and genecompared. ResultsAfter 4 times of screening, abundant of specific phage modeling peptides were harvested. The results of ELISA showed a fine associativity of some phage modeling peptides. A total of 11 sequences were recognized which were related to promoting cell division and proliferation or inhibiting cell proliferation. ConclusionhFGF7 related phage modeling peptides can be harvested from a phage random twelve peptide library， in which some phage modeling peptides may promote epidermis cells proliferation.

Effects of pirfenidone on liver fibrosis in rats

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  50-54. 

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To investigate the effects of pirfenidone (PFD) on liver fibrosis in experimental rats. MethodsFortyfive male Wistar rats were randomly divided into three groups. The model group was intraperitoneally injected with 1% dimethylnitrosamine (DMN) 10?mg/kg per day for the first three days of each week for 4 weeks. The intervention group was orally given PFD 500?mg/kg every day for 4 weeks starting on the day of the first injection of DMN. The control group was given normal sodium. Rats were killed at the end of the 4th week, and the livers and spleens were removed. Blood samples were obtained immediately before rats were killed. Liver histological and ultramicroscopic changes were observed, serum indices of liver fibrosis and liver function were examined, and the level of transforming growth factor β1 (TGFβ1) was determined. ResultsCompared with the model group, the intervention group had a higher bodyweight(P＜0.05) but splenohepatomegalia was less(P＜0.05). Serum levels of hyaluronic acid, laminin and type Ⅳ collagen of the intervention group were significantly lower than those of the model group(P＜0.05), however serum levels of alanine aminotransferase(ALT), aspartate aminotransferase (AST), total bilirubin（TBIL） and TGFβ1 were significantly higher than those of the model group(P＜0.001). Masson trichrome staining showed that the intervention group had mild bridging fibrosis and less lymphocyte infiltration. No typical pseudolobule was found. Transmission electron microscopy demonstrated that hepatic cell injury in the intervention group was significantly less than that in the model group and slight collagen fibers were observed in the sinus hepaticus and Disse space. ConclusionsPFD has a significant antifibrosis effect in experimental rats, which was presumably caused by inhibiting the production of TGFβ1.

Effect of nuclear factorκB inhibitor PDTC on pulmonary arterial hypertension

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  55-58. 

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To investigate changes and functions of nuclear factor κB (NF κB) and its inhibitor PDTC in monocrotaline（MCT）induced pulmonary arterial hypertension（PAH）. MethodsPulmonary arterial hypertension rat models were established by injecting MCT toxin. A total of 58 Wistar rats were randomly divided into 7 groups : the MCT0 group, the MCT1W group, the MCT2W group, the MCT3W group, the control/vehicle group, the MCT/ vehicle group, and the MCT/PDTC group. Hemodynamic studies were determined by right cardiac catheterization, activity of NFκB was determined by EMSA, and expression of intercellular adhesion molecule1 (ICAM1) and macrophage infiltration were determined by immunohistochemistry. ResultsICAM1 expression and macrophage infiltration were significantly upregulated from week 1 after injection(P<0.01), and the mean  right ventricular pressure and indexes of right ventricular hypertrophy increased from week 2 after injection(P<0.01). Compared with controls, MCT treatment increased the mean  right ventricular pressure (27.8±1.5?mmHg  vs 17.2±1.4?mmHg, P<0.01), which was reduced by PDTC treatment(18.4±2.2?mmHg, P<0.01). Indexes of right ventricular hypertrophy induced by MCT were similarly inhibited (P<0.01) by PDTC treatment, which was associated with suppression of ICAM1 expression and macrophage infiltration. ConclusionWe conclude that NFκB activity and ICAM1 expression are probably associated with the development of MCTinduced PAH, which is ameliorated by administering a NFκB inhibitor, PDTC.

Human fetal liver cell transplantation in treatment of mucopolysaccharidosis

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  59-63. 

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ObjectiveTo study the immunological reaction of mice with mucopolysaccharidosis treated by human fetal liver cell transplantation in order to select an  immunosuppressive agent. MethodHuman fetal liver cells(FLCs) were transplanted into αLduronidase (IDUA) deficiency mice pretreated with different immunosuppressive agents. T cell subsets（CD3,CD4/CD8）were detected by flowcytometry, while cytokines(IgG,IgM,IL2, TNFα and IFNγ) by EL ISA, and serum  IDUA activity by a fluorospectrophotometer at different stages (before and after transplantation). ResultsThe T cell subsets and cytokines in recipients were significantly increased after FLCs transplantation. Five days after transplantation, IDUA activity reached a peak in all groups (P＜0.01), then gradually decreased. The IDUA activity lasted the longest in the group pretreated with cyclophosphamide cyclosporine FTY720 and predinisone. ConclusionIDUA activity can be raised by FLCs transplantation. Due to immunological rejection, the recipient should be given immunosuppressive therapy. Cyclophosphamide cyclosporine FTY720 in combination with predinisone can give the best immunosuppression.

Specific siRNA inhibits HPV16 E7 gene expression: an experimental study

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  64-67. 

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To screen an effective HPV16 E7 specific siRNA and investigate its effect on expression of HPV16 E7 mRNA and cell surface HLA class I antigen in HaCaTE7 cells. MethodsThree HPV16 E7 specific siRNAs were chemically synthesized and transfected into HaCaTE7 cells by Lipofectamine2000. Expression of HPV16 E7 mRNA was determined by realtime PCR while expression of cell surface HLA class I antigen was determined by flow cytometry（FCM）.  ResultsAll three siRNAs effectively inhibited expression of HPV16 E7 mRNA in HcaCaTE7 cells. Of the siRNAs, siRNA2 was the most effective and its inhibitory rate reached 75%. Expression of cell surface HLA class I antigen was obviously upregulated. Posttransfected average mean fluorescence intensity (MFI) of HaCaTE7 cell surface HLA class I antigen was 130.18±1.07, which was significantly higher than that in the mock control group (100.32±3.01) and the nonspecific siRNA control group (100.82±2.87). ConclusionHPV16 E7 specific siRNA can effectively inhibit expression of

Effects of the Rosiglitazone on expressions of MMP2 and TIMP1 mRNA in hepatic stellate cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  74-75. 

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To investigate the effects of Rosiglitazone on expressions of MMP2 and TIMP1 mRNA in hepatic stellate cells (HSCs) in vitro. MethodsThe HSC line, HSCT6 was incubated with different concentrations of Rosiglitazone. The MMP2 and TIMP1 mRNA levels were measured by reverse transcription polymerase chain reaction (RTPCR).  ResultsExpression of TIMP1 mRNA in the Rosiglitazonetreatment groups was significantly lower than that in the control group (P＜0.05), and there was no difference in the MMP2 mRNA levels between the Rosiglitazonetreatment groups and the control group. ConclusionThe antifibrosis mechanism of Rosiglitazone is partly due to its downregulation on TIMP1 but not on MMP2 expression of hepatic stellate cells.

Effects of the SPK/S1P signal on proliferation and apoptosis of human hepatoma cell HepG2

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  76-79. 

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To investigate the effect of the SPK/S1P signal on proliferation and  apoptosis in human hepatoma cells. MethodsTreated with 520?μmol/L dimethyl sphingosine (DMS) in culture medium, human hepatoma cell HepG2 was determined using MTT assay, a scanning electron microscope and TUNEL staining. ResultsMTT assay demonstrated that DMS inhibited the growth of human hepatocarcinoma HepG2 cells in a timeand dosedependent manner. HepG2 cells showed typical morphological characters of apoptosis. TUNEL demonstrated that the stained cells of the DMS group were much more than those of the control group, which had significant differences(P＜0.01）. ConclusionDMS inhibits the growth of human hepatocarcinoma HepG2 cells, and can induce apoptosis of human  hepatocarcinoma HepG2 cells.

Relationship between DDR2 and alcoholic liver fibrosis and response of compound glycyrrhizin tablets on them: an experimental study

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  80-84. 

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To investigate the relationship between DDR2 and alcoholic liver fibrosis as well as the response to compound glycyrrhizin tablets treatment on them. Methods48 rats were randomly divided into 6 groups (8 in each group): the control group, the model group, the Alc disc group, the Alc contin group, the CGT + Alc disc group and the CGT + Alc contin group. After 16week intragastric administration with alcohol, rats in the model group were sacrificed, at the same time rats in the control group were killed. The rest having been treated by intrastric alcoholic for 16 weeks in the Alc disc group (stopped alcohol intragastric), the CGT + Alc disc group (stopped alcohol intragastric and treated by CGT), the Alc contin group (continued alcohol intragastric without treatment of CGT) and the CGT + Alc contin group (continued alcohol intragastric with treatment of CGT simultaneously) were sacrificed at the end of the 20th week. Levels of serum hyaluronic acid (HA), laminin (LN), procollagen type Ⅲ (PCⅢ) and collagen type Ⅳ (CⅣ) were determined. Pathologic changes of the liver were observed by HE staining and Masson staining. Expressions of DDR2 mRNA and protein were determined by RTPCR and Western blot, respectively. ResultsPathological liver fibrosis was obviously found in rats with alcoholic intragastric administration for 16 weeks. Areadensity percentage of collagen fibrosis, levels of serum HA, LN, PCⅢ and CⅣ, and levels of DDR2 mRNA and protein in the liver were significantly increased in these indexes were decreased in the CGT + Alc disc group compared with the Alc disc group and in the CGT + Alc contin group compared with the Alc contin group(P＜0.05). ConclusionCGT had certain therapeutic effects on alcoholic liver diseases, which may relate to downregulation of DDR2 expression.

Monocytes were induced to express the markers of lymphatic endothelial cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  85-88. 

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Lymphangiogenesis is thought to be involved in the pathophysiological process such as tumors and inflammation. This research looked into the possibility of the transdifferentiation of mononuclearmacrophages into lymphatic endothelial cells in inflammatory conditions. MethodsFresh monocytes from peripheral blood were collected and cultured in ECM. The cells were next incubated in FNplated wells or treated with VEGFC, TNFα and LPS for 24 hours respectively. Expression of specific markers of lymphatic endothelial cells such as LYVE1, Podoplanin and Prox1, and the common markers of endothelial cells such as vWF and eNOS were detected by RTPCR and immunochemical methods. ResultsBefore the induction,  expression of LYVE1 was  positively detected  in cultured mononuclearmacrophages, while the expression of vWF, eNOS, Podoplanin, Prox1 was negatively detected. After the induction, the level of expression of Podoplanin and Prox1 obviously increased in cultured mononuclearmacrophages, yet expression of vWF and eNOS remained negative. ConclusionMononuclearmacrophages can be induced to express the markers of lymphatic endothelial cells by FN, VEGFC, TNFα and LPS.

Comparison of different sample preparation methods for human serum 2DE

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  89-94. 

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To compare different sample preparation methods for human serum 2DE. MethodsSerum samples were prepared by five different methods: acetone precipitation, trichloroacetic acid （TCA） precipitation, acetone /TCA precipitation, 2D clean up kit, and Albumin and IgG Removal Kit. Then, after the samples were run on two dimensional gel electrophoresis (2DE), 2DE gels were stained and analyzed by Image Master 5.0, and protein spots were identified by MALDITOF MS. ResultsThe total protein number of samples processed by acetone precipitation, TCA precipitation, Acetone/TCA precipitation, and 2D clean up kit was 1?802±12, 1?648±10, 1?727±13 and 1?670±14, respectively. In addition, the total protein number of samples, one of which was processed by sample preparation kits removing the albumin and immunity goblin, the other one of which was not, was 1?212±16 and 1?258±12, respectively. Conclusion2DE maps obtained from samples treated by different serum precipitation methods were different. By removing albumin and immunity goblin from the serum, low abundance proteins were increased, but some high abundance proteins were lost.

Differentiation and expansion of megakaryocytes from CD34+ cells derived from umbilical cord blood and change of gene expression

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  95-99. 

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To investigate the differentiation and expansion of megakaryocytes from CD34+ cells of umbilical cord blood and observe the change of their specific gene expression. MethodsCD34+ cells were obtained by  the immunomagnetic bead method and cultured in serumfree and stromafree media containing the following three cytokine combinations: thrombopoietin (TPO) +stem cell factor(SCF)+interleukin(IL)3+IL6,TPO+SCF+IL3 and TPO+SCF. The cells were collected on the 3rd, 7th, 10th and 14th day and expression of cell surface molecules was determined by fluorescence microscopy, and then cell apoptosis was determined by flow cytometery(FCM), maturation evaluation of the Mk ploidy was also carried out, and specific gene expression was determined by real time fluorescent quantitation PCR. ResultsIn vitro the CD34+ cells were effectively expanded. With the increase of time, expressions of CD41+, CD42b+, and CD61+ cells did not significantly change, but the optimal expansion of the Mk progenitors(CD34+/CD41+) was observed at day 7. Degree of maturation of the Mk cells also increased by evaluating the DNA content. TPO+SCF+IL3+IL6 was the optimal expansion method. Realtime fluorescent quantitation PCR displayed that transcription factor GATA1, NFE2, TXS, GPIIb and HPRT reached high levels at the 7th day, but

Effects of Lipopolysaccharide on the proliferation and apoptosis of human dental pulp cells

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  100-102. 

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To observe lipopolysaccharide(LPS) induced apoptosis of human dental pulp cells(HDPC) and its effect on apoptosisrelated cystine protease 3 (caspase3), Bcl2, and Bax activity in vitro. MethodsPulp cells were stimulated with 0, 0.01, 0.10, 1.00, 10.00, 100.00?mg/L LPS. Then the cell proliferation activity was detected using MTT colorimetrics, the cell cycle and apoptosis after LPS stimulation using flow cytometry, and expressions of caspase3, Bcl2, Bax using immunocytochemical staining. ResultsCell growth was significantly inhibited by LPS in a dosedependent manner.  Flow cytometry results showed that  the apoptotic rates were respectively 3.1%, 3.5%, 12.1%, 14.4%, 24.3%, and 59.7% after stimulated by LPS. Immunocytochemical staining showed that Bax expression was increased, while caspase3 and Bcl2 expressions were decreased. ConclusionLPS could significantly inhibit the proliferation of HDPC, induces apoptosis, reduces Bcl2 and caspase3 activity, and increases Bax activity, which is dosedependent.

Role of apolipoprotein E gene polymorphism and plasma lipids in mild cognitive impairment

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  103-107. 

Abstract ( 1063 )   PDF (361KB) ( 228 )   Save

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To compare the distribution of Apolipoprotein E(ApoE) genotypes and plasma lipid levels between patients with mild cognitive impairment (MCI) and normal healthy people. Methods120 patients with MCI and 120 normal controls were examined with a neuropsychological test which included minimental state examination (MMSE), activities of daily living scale (ADL), global deterioration scale ( GDS) and Hachinski ischaemic scale (HIS). Blood TC, TG, HDLC and LDLC were determined, and the ApoE phenotypes and allele frequency were determined using PCRRFLP. ResultsMCI cases achieved significantly lower scores than normal controls in memory, execution function, attention, calculation, and directive force. Between the MCI and the control groups there was a statistical difference in ApoE genotypes and allele frequency. The ApoEε3/4 genotype and ApoEε4 in the MCI group were higher than those in the normal control (P＜0.05). TC and LDLC in the MCI group was higher than those in the normal controls (P＜0.05). In the MCI and control groups, TC and LDLC levels of ApoE ε4 were higher than those of ApoE ε3 and ApoE ε2（P＜0.05）. ConclusionApoE genotypes and allele frequency are statistically different between the MCI and the control groups.  ApoEε4 allele frequency is a risk factor of MCI. Dyslipidemia is found in MCI.

Expression and methylation of p16 in carcinoma ex pleomorphic adenoma of the parotid gland

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  108-110. 

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To study the changes of p16 gene protein expression and promoter methylation in carcinoma ex pleomorphic adenoma (CXPA) of the parotid gland, and to evaluate their significance for carcinogenesis of CXPA.  Methodsp16 gene protein expression and promoter methylation were determined in 40 samples of CXPAs of the parotid glands and 10 normal samples of the parotid gland(NM) using immunohistochemistry (IHC) and methylation specific PCR (MSP).  ResultsIn normal parotid glands, the p16 protein was positively expressed. In the CXPAs group, 21 cases (52.5%) showed a decreased (in different degrees) or absence of p16 protein expression. There was significant statistical difference (P＜0.05) between the two groups. Hypermethylation of the p16 gene promoter was found in 16 cases (16/40, 40%) of CXPAs, but was not found in normal salivary glands (P＜0.05). ConclusionThe decreased or absence of p16 protein expression and promoter hypermethylation play an important role in the carcinogenesis of CXPA. And hypermethylation may be the important inactivation mechanism of the p16 gene in CXPA.

Effect of Coptis decoction on gingivitis in fixed orthodontic treatment

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  111-113. 

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To investigate the effect of Coptis decoction on gingivitis in fixed orthodontic treatment. Methods100 patients aged 11 to 19 were randomly divided into experimental group (50 patients) and control group (50 patients). The experimental group was treated with Coptis decoction besides normal oral health measures, while the control group was treated with 0.9% N.S besides normal oral health measures. Changes of GI (gingival index) and PLD (plaque index) in the second premolar and first molar of the two groups were measured before and after the treatment. Results4 weeks after the fixed appliance was placed, GI and PLI were increased in both groups, but there was no significant difference between the two groups, however 8 weeks and 12 weeks later, GI and PLI in the two groups showed significant differences (P＜0.05). ConclusionCoptis decoction can efficiently reduce the incidence of gingivitis in fixed orthodontic treatment.

Quantitative analysis of cardiac troponin T and the risk factors of myocardial infarction

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  114-117. 

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To investigate the effect of incidence of acute myocardial infarction (AMI) associated with diabetes mellitus or hypertension on myocardial infarction size.  Methods62 patients with AMI hospitalized from June 2006 to June 2008 were divided into three groups according to complications: group A contained 18 patients who had AMI with diabetes mellitus, group B contained 21 patients who had AMI with hypertension, and group C contained 23 patients who had AMI without diabetes mellitus or hypertension. Then cTnT, CKMBmass and CKMBact were kinetically measured in these patients. Results①Abnormal changes of serum cTnT, CKMBmass, and CKMBact levels appeared at 3.9, 2.9, 3.0?h respectively in group A, while at 3.5, 1.6, 2.3?h in group B, and at 2.8, 1.3, 1.7?h respectively in group C. ② The maximal values of cTnT, CKMBmass and CKMBact appeared at 38.9, 28.5, 32.4?h in group A, at 35.4, 26.7, 29.5?h in group B, and at 30.5, 18.2, 20.7?h in group C. ③ cTnT, CKMBmass and CKMBact returned to the baseline levels at 78.5, 72.6, 63.2?h in group A, at 58.7, 52.4, 41.7?h in group B, and at 62.5, 58.6, 52.5?h in group C. ④ Quantity of injured myocardial in group A was 45.2 cTnTg, 42.5 CKMBmassg, and 45.7 CKMBactg; in group B was 40.8 cTnTg, 37.4 CKMBmassg, and 39.2 CKMBactg; ingroup C was 31.4 cTnTg, 28.7 CKMBmassg, and 31.6 CKMBactg. ConclusionsAMI patients complicated with diabetes mellitus and hypertension have highly critical conditions, which maybe attribute to an increased myocardial infarct size.

31phosphorus magnetic resonance spectroscopy in  diagnosis of the benign and malignant masses in liver

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  118-120. 

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ObjectiveTo probe the clinical value of 31phosphorus magnetic resonance spectroscopy(31PMRS) in vivo at 3.0 Tesla in diagnosis of the benign and malignant masses. Methods31PMRS scanning with a single voxel was carried out on 35 benign tumors, 62 malignant ones and 13 hepatic abscesses. Intracellular pH value (pHi), and some metabolic parameters including phosphomonoester (PME), phosphodiester (PDE), inorganic phosphate (Pi), γ, β, and α adenosine triphosphate (ATP), lower energetic phosphate (LEP), and the ratios of PME/ATP, Pi/ATP, PME/PDE, PME/Pi, PDE/Pi and PDE/ATP were calculated. The differences in the metabolic parameters above between benign and malignant groups were analyzed. ResultsThe differences inγATP, which were 1?279.46±432.21 and 758.22±240.79 in benign and malignant lesions respectively, and Pi/ATP, 3.98±0.8 and 5.36±0.96, were identified statistically (P＜0.05), and so were the differences in pHi, 7.03±0.43，7.09±0.35 and 7.45±0.51 in benign, malignant tumor and abscesses respectively, and γATP, 1?260.71±407.72，758.22±240.79 and 1?268.62±444.10(P＜0.05). However, differences in other metabolic parameters among these groups were not found statistically (P＞0.05). ConclusionSuch phosphorus metabolites as γATP and Pi/ATP in HCC may be used to differentiate the benign and malignant masses, while pHi is helpful for the diagnosis of hepatic abscesses.

Surgical treatment of aortopulmonary septal defect

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  121-123. 

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To summarize experience in surgical treatment of aortopulmonary septal defect(APSD). Methods15 patients with APSD were operated from October 2000 to January 2009. They were divided based on the Richardson′s classification: typeⅠ 8 cases, typeⅡ 5 cases, and type Ⅲ 2 cases. Among typeⅠ, 2 cases received Dacron repair across of the frontal septal defect. Among type Ⅲ, 1 case underwent the treatment with an artificial conduit connecting the RPA with MPA, another received the treatment of connecting the RPA directly with MPA. Others received Dacron repair across of the aorta. ResultsIn the whole group,there was no operative death. 8 patients had clinical improvement at a followup of 5 months to 3 years. There was no shunting, aorta and pulmonary artery stenosis. Pulmonary artery pressure obviously decreased. ConclusionThis operation is suitable for young infants so as to avoid pulmonary vascular disease, and preoperative accurate diagnosis and full understanding of pathophysiology and clinical anatomy are the keys for success.

Uncertainty of the content of Glibenclamide in Chinese Traditional Medicine determined by HPLC

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  124-127. 

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To establish a method for evaluation of uncertainty of determination of glibenclamide illegally added in Jiangtangning by HPLC. MethodsThe uncertainty was evaluated by a established mathematical model and various factors affecting sample detection were analyzed. ResultsUncertainty of each componet was calculated and their combined uncertainty was then finally obtained by synthesizing the uncertainties of various component variables. ConclusionThis method is appropriate to be used in uncertainty evaluation for determination of declofenac sodium in Chinese traditional medicine by HPLC.

Mixed micelles made of Pluronic P105 and L61 as pharmaceutical  nanocarriers for camptothecin

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  128-131. 

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To increase the drug loading of the poorly soluble anticancer drug, camptothecin (CPT)， micelles made of a mixture of Pluronic P105 (P105) and Pluronic L61 (L61) were prepared. MethodsMixed micelles made of a mixture of Pluronic P105 (P105) and Pluronic L61 (L61) were prepared loaded with CPT. The drug loading and colloidal stability of such systems were studied, and cytotoxicity of the CPTloaded mixed micelles against MCF7 cancer cells in vitro was determined. ResultsThe CPTloaded mixed micelles were stable upon storage and dilution. Drug loading of CPT was increased with an increase of the amount of Pluronic L61. Cytotoxicity of the CPTloaded mixed micelles against MCF7 cancer cells in vitro was remarkably higher than that of the free drug. ConclusionMixed micelles made of a mixture of Pluronic P105 (P105) and Pluronic L61 (L61) may serve as a stable delivery system for poorly soluble CPT.

China college student personality scale and adjustment scale test for 6913 freshmen

JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES). 2009, 47(11):  132-134. 

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To explore the correlation between personality and adaptability of freshmen. MethodsUsing the China College Student Adjustment Scale (CCSAS) and China College Student Personality Scale (CCSPS), we undertook a survey about mental health for freshmen at a university in Shandong Province and then analyzed it with Pearson linear correlation analysis and canonical correlation analysis. ResultsMany factors between CCSAS and CCSPS had positive correlations. The active factor, tenacity factor, and easygoing factor in CCSPS were strongly related to factors of CCSAS. The first typical correlation coefficient was 0.844?1 and the  eigenvalue contribution rate was 76.5%. The mental adaptation from interpersonal and self relationships is correlated with all factors in CCSPS.  ConclusionFactors of CCSAS are statistically significantly related to factors of CCSPS.