Abstract:Objective
To prepare the anti-Listeria monocytogenes(LM) polyclonal antibody derived from rabbit and to study the application of LM in latex agglutination test(LAT).Methods
Standard LM strain were cultured and multiplied using selective agents and enrichment  procedures.Freund’s complete inactivated vaccine and Freund’s incomplete inactivated vaccine were made for immunizing rabbits.Blood samples were collected from the marginal ear vein and the antibody titers were measured by ELISA.Four times after immunization,blood serum was separated from the carotid artery and crude IgG was purified  by using the method of caprylic acid-sulfide saturated soution precipitation.IgG molecules were purified by using the HiTrap Protein G chromatography column on AKTA Protein purification device and identified using the methods of SDS-PAGE electrophoresis,UV spectrophotometry,and indirect ELISA.Purified IgG were covalent conjuncted with latex microspheres and the content of latex and interaction time were optimized.The method was established for rapid detection of LM in simulated samples.Results The protein content of the purified anti-LM IgG was 23.364 g·L^-1 and its antibody titer was 1∶12　800-1∶256 00,no cross-reaction occured with a variety of bacteria such as Vibrio cholerae,Salmonella,Shigella,but a weak cross-reaction with Steinmann Listeria monocytogenes.The covalent conjuncted ratio of IgG on the latex microspheres was 1∶40 and the positive rate was 87.5% for detetion in 246 samples by the method of LAT.The minimum detectable content of the antigen was 0.039 8 g·L^-1.Conclusion LAT is a promising method to facilitate the grassroots level for field testing LM with its specificity,high sensitivity and good reproducibility.

Abstract:Objective
 To expiore the influence of  oral low dose of mixed rare earth　Changle in the  morphological structure of the kidney of diabetic rats, and clarify its repair on  kidney structure damage of diabetic rats.Methods  Male Wistar rats were treated with STZ to set up diabetes mellitus models and then  divided into  diabetic model group and Changle treatment group (n=10),the other 10 nomal rats were used as control group,the rats in three groups were given daily saline, Changle(0.2 mg?kg-1),and daily saline,orally one month, then they were executed and the kidney was obtained.The　kidney morphometry was used to detect the changes  of  rat kidney weight, body weight
, kidney weight to body weight ratio, renal cortical thickness, glomerular volume density, total glomerular volume, near (far) end of the total volume, near (far) end of the tubular epithelial cell height, near (far) end of tube cavity diameter  and the morphological changes of kidney.Results  Compared with control group and Changle treatment group,in diabetic model group, the kidney weight was increased significantly(P<0.01), the body weight was markedly decreased (P<0.01),the kidney weight to body weight ratio was significantly increased(P<0.01);the total glomerular volume, near (far) end of a small tube volume, proximal tubule epithelial cell height, near (far) end of  the small tube cavity diameter were significantly increased  (all P<0.01); the kidney cortical thickness, glomerular volume and diatal tubule epithelial cells had no significant change(P>0.05).Compared with control group, in Changle treatment group the kidney weight, renal cortical thickness and glomerular volume density, glomerular total volume, total vovlume of proximal tubule, near (far) end of the tubular epithelial cell height, near (far) end of tube cavity diameter had no significant difference(P>0.05), but the body weight was significantly reduced (P<0.01), and the kidney weight to body weight  ratio,near (far) end of a small tube,and  total volume were significantly increased  (P<0.01).The HE staining results showed that the kidneys in control  and Changle treatment groups were normal, the rat kidney in diabetic model group was swollen;the  PAS staining results showed that in diabetic model group most of the proximal tubular epithelial cells were markedly swollen, containing increased PAS positive material, a small number of  proximal tubules showed the shedding of epithelial cells, lumen expansion and part of expansion of the distal tubule; however there were no significant changes in epithelial cells;in normal control group and Changle treatment group there were no the  changes mentioned above.Conclusion Oral low dose of  mixed rare earth Changle can repair  structure damage of  the kidney of the  diabetic rats.

bstract:Objective
To observe the effects of the intrathecal injection of N-methyl-D-aspartate (NMDA) receptor agonist NMDA and antagonist MK801 on the cardiacsomatic motor reflex (CMR) induced by intrapericardial  bradykinin (BK), and explore the role of Glu receptor NMDA subunit in nociceptive information transmission of the heart in the spinal cord.
Methods 26 male SD rats were randomly divided into BK group(n=8), BK + NMDA group(n=6), BK + MK801 group(n=6), and BK + MK801 + NMDA group(n=6).CMR was induced by intrapericardial  BK, and the effects of intrathecal injection of drug in various groups on CMR were observed.The changes of CMR were monitored via electromyogram (EMG) responses of dorsal spinotrapezius muscle to intrapericardial  BK.Results There
 were no significant differences among EMG induced by intrapericardial  BK, four times at 40 min intervals.Compared with control, the EMG was increased from 100% of control to (149.86±8.54)% by intrathecal injection of NMDA, there was significant difference of EMG between before and after intrathecal injection of NMDA (P<0.05).However, the EMG was decreased from 100% of control to (96.22±2.31)% by intrathecal injection of MK801, there was  no statistically significant difference (P>0.05).The EMG was increased from 100% of control to (103.09±4.13)% by intrathecal combinative injection of MK801 and NMDA, there was no significant difference of EMG between before and after intrathecal combined injection of MK801 and NMDA (P>0.05).Conclusion CMR induced by intrapericardial  BK has reliable repeatability and the spinal cord level of NMDA receptor involves in the modulation of CMR.

Abstract:Objective
To study the influence of 17β-estradiol and progestogen in type Ⅱalveolar epithelial Na+ channel（ENaC）in rats with acute lung injury(ALI) and explore the mechanism of estradiol and progestogen in  promoting the  clearance of alveolar fluid.Methods 40 immature female SD rats were randomly assigned into 4 groups: normal control group(injected with 8 mL?kg-1 saline via venous),ALI model group,17β-estradiol and  progestogen  combination A and B group.The rats in last three group were injected with 0.1  mL?kg-1 oleic acid via venous to establish ALI model.The rats in combination A and B groups  were injected subcutaneously with 17β-estradiol and  progestogen mixed liquor 0.1 mL in different ratios (A group 1∶1　000，B group 1∶200) at 12 and 36 h before oleic acid injection.All the rats were sacrificed at 6 h after oleic acid injection.The wet/dry ratio and the lung coefficient were determined, the pathological changes of  lung  tissues  were  observed, and the expressions of α-ENaC,β-ENaC and γ-ENaC mRNA and protein were determined.Results
① Compared with normal control groups,the wet/dry ratio and the lung coefficient in ALI model group were increased significantly (P<0.01);in combination A and B groups, the wet/dry ratios and the lung coefficients were decreased compared with model group，especially in  A group (P<0.05).②Obvious pulmonary edema,bleeding and inflammatory cell infiltration were observed in ALI group.However, the extent of  pulmonary edema,bleeding and inflammatory cell infiltration were decreased markedly in combination  A and B groups, especially in  A groups.③ Compared with normal control group,the  expressions of α-ENaC and γ-ENaC mRNA and protein were decreased in ALI model group(P<0.01); compared with ALI group,they were increased significantly in combination A and B groups,especially in A  group(P<0.05).Conclusion 17β-estradiol and progestogen can increase the expressions of  type Ⅱ alveolar epithelial α-ENaC and γ-ENaC mRNA and protein in rats with ALI,and promote the clearance of alveolar fluid.

Abstract:Objective
To study the role of p16 in cigarette smoking-induced lung carcinogenesis by detecting the expression of p16 in human normal bronchial epithelial cells（HBE） after exposure to cigarette smoke extracts (CSE) and analyze the underlying mechanism.Methods HBE were treated with various doses of CSE or control DMSO and the protein and mRNA levels of p16 were determined by Western blotting and Realtime RT-PCR respectively.The genomic promoter of p16 was cloned and its transcriptional activity after exposure to CSE was determined by luciferase assay.Results  After the treatment of different doses (0,10,25,50,100 mg•L^-1) of CSE,the mRNA and protein levels in HBE were significantly inhibited in a dose-dependent manner.More than 25 mg•L^-1 of CSE conld significantly inhibit the  p16 mRNA and protein levels,there was significant difference compared with control group(P<0.05).100 mg•L^-1 of CSE showed the greatest inhibiory effect. Moreover,the transcriptional activity of the promoter of p16 was significantly suppressed by more than 25 mg•L^-1 CSE,there was significant difference compared with control group(P<0.05).Conclusion Cigarette smoke extracts can inhibit the expression of p16 by suppressing its transcriptional activity,indicating that p16 might play a key role in cigarette smoking-induced lung carcinogenesis.

Abstract:Objective
 To explore the development change of Kv1.3 and Kir2.1 channels in human macrophages and the effects of gemfibrozil on both channels’ expression and their membrane potentials,which will provide electrophysiologic basis for its treating atherosclerotic diseases.Methods The obtained human monocyte-derived macrophages were randomly divided into 3 groups: control 5 d group (C 5d),control 7.5 d group (C 7.5d) and gemfibrozil group (G).The final　concentration of gemfibrozil was 400  μmo•L^-1.The effect of gemfibrozil on the expressions of Kv1.3 and Kir2.1 channels was investigated using Real time RT-PCR and Western blotting,and its effect on membrane potentials was analyzed with the optical mapping of the membrane potential with the voltage-sensitive dyes. Results  Compared with  C 5d group,the expressions of Kv1.3 mRNA/protein in  C 7.5d group were raised from (1.064±0.275)/(0.227±0.018) to (3.067±0.824)/(0.409±0.022) (P<0.05),but the expressions of Kir2.1 mRNA/protein were decreased from (1.024±0.166)/(0.204±0.018) to (0.399±0.133)/(0.042±0.008)（P<0.05）. Compared with  C 7.5d group,the expressions of Kv1.3 mRNA/protein in  gemfibrozil group were reduced to (1.137±0.067)/(0.143±0.023) (P<0.01); however,the expressions of Kir2.1 mRNA/protein were elevated to (1.35±0.087)/(0.202±0.033)（P<0.01）.Meanwhile,gemfibrozil significantly reduced the surface fluorescence intensity of the macrophages,and the membrane potentials in  C 5d and C 7.5d groups were decreased from (1.000±0.026) to (0.833±0.046) and (0.481±0.053),respectively (P<0.05).Conclusion Gemfibrozil differentially regulates the expressions of Kv1.3 and Kir2.1 channels in human monocyte-derived macrophages,
and lowered their membrane potentials.

Abstract:Objective
To investigate the effects of photodynamic therapy mediated by meso-5, 10, 15, 20-Tetrakis-(N-methyl-4-pyridyl)porphine(TMPyP4) on human ovarian carcinoma A2780 cells in vitro as well as the expression levels of MCM2 and CA-Ⅸ  mRNA in A2780 cells and to clarify the destruction mechanism of the TMPyP4-PDT on human ovarian carcinoma A2780 cells.Methods The human ovarian carcinoma A2780 cells were cultured in vitro and divided into 4 groups as follows: blank control group, simple TMPyP4 group, simple PDT group and TMPyP4-PDT group.The  increase of inhibitory effects of TMPyP4-PDT on proliferation of A2780 cells were detected by CCK-8 assay.The apoptosis of A2780 cells after treated with different concentrations of TMPyP4 were detected by flow cytometry(FCM).The morphological changes of cells in various groups were observed with HE staining under light microscope.The expression levels of MCM2 and CA-Ⅸ  mRNA were detected by RT-PCR.Results  The  inhibitory rates of cell proliferation in simple TMPyP4 group were gradually increased with the increase of the dosage(P<0.05).The increase of inhibitory rates of cell proliferation in simple PDT group was not obvious(P> 0.05).The inhibitory rates of cell proliferation in TMPyP4-PDT group were increased with the increase of the dosage and the light energy density(P<0.05).FCM analysis showed that TMPyP4-PDT could induce the apoptosis of A2780 cells; the apoptotic rates of A2780 cells were (9.46±0.04)%, (17.7±0.3)%, (25.4±0.2)%, (30.2±0.2)% and (66.3±0.3)% when the light energy density was 4 J•cm^-2 and the doses of TMPyP4 were 3, 6, 15, 30 and 60 μmol•L^-1 and the treatment time was 4 h. More adherent cells without nuclear pyknosis and nuclear fragmentation in blank control group and simple PDT group were observed under light microscope; Less adherent cells as well as nuclear pyknosis, vacuole and some cytoplasmic dissolution in simple TMPyP4 group and TMPyP4-PDT group could be observed under light microscope.RT-PCR analysis showed that compared with blank control group , the expressions of MCM2 and　CA-Ⅸ  mRNA were decreased significantly after treated with 3, 6, 15, 30 and 60 μmol?L-1 TMPyP4 for 4 h (P<0.01) under the conditions of the light energy density of 4 J?cm-2.Conclusion TMPyP4-PDT has distinctive inhibitory effects on human ovarian carcinoma A2780 cells, and the mechanism may associate with the negative regulation of the expressions of MCM2 and CA-Ⅸ .

Abstract:Objective
To study the long-term effects of lasting and high-dose  electromagnetic radiation on number and percent of peripheral immune cells in mice and explore the influence of the lasting and high-dose electromagnetic radiation in murine immune system.Methods  Thirty BALB/c mice were randomly divided into control group,CTX group and irradiation group.The mice in irradiation group were exposed to electromagnetic radiation at the power density of 10 mW•cm^-2,30 min a day,for 5 consecutive days every week,lasting for 4 weeks.The mice in CTX group were fed with CTX with the concentration of 30　mg•kg^-1,for seven times.The number and percents of CD4+T cells,CD8+T cells,CD4+/CD8+,CD49+ NK cells in peripheral blood cells were detected 30,45,60,75,90,105 and 120 d after electromagnetic radiation.Results After high-dose electromagnetic radiation,the number of WBC was increased obviously (P<0.05).The number of CD4+ T cells was increased on the 75th day (P<0.05) and began to decrease on the 90th day,while the number was much more than that in control group (0 mW•cm^-2).Until the 120th day,the number of T cells was as the same as the control.After the long-term effect of lasting and high-dose electromagnetic radiation,the number of CD8+T cells was decreased,and from the 90th day to the 120th day the number　of T cells became normal (P<0.05).The ratio of CD4+/CD8+ and the number of CD49+ NK cells were lower than those in control group after irradiation (P<0.05).
Conclusion Chronic,persistent large doses of electromagnetic radiation can lead to immune system disorders,through detecting the  changes in the number of peripheral blood CD4+ T cells,CD8+ T cells,CD49+ NK cells and the proportion of CD4+/CD8+,the immune status of mice after irradiation can be found.

Abstract:Objective
To build oxygen glucose deprivation/reperfusion(OGD/R) model in vitro to mimic ischemia and reperfusion inducing apoptosis of SH-SY5Y cells,to detect differential proteins by employing proteomics technique and to reveal the potential   mechanisms of neuronal apoptosis.Methods  SH-SY5Y cells were employed and divided into control group and experimental group in which the cells treated with OGD10h/R24h to set up apoptosis model.After the proteins were extracted from SH-SY5Y cells in control and OGD10h/R24h groups,2D-DIGE techniques was used to detect differential proteins.Results  By DeCyder software mean (1 800±156) protein spots were  displayed on 2D-DIGE image.In OGD10h/R24h group,30 protein spots were detected to  differentially express compared with control group,of which 22 spots were up-regulated and 8 spots were down-regulated.Compared with control groups at the same time, the differences were significant (P<0.05 or P<0.01).Conclusion  By employing 2D-DIGE proteomics techniques to analyse OGD/R model,the protein spots are detected to differentially express.

Abstract:Objective
 To study the proteomic differences between malignant melanoma tissue and normal skin tissue and to look for the possible marker for diagnosis and therapy target of malignant melanoma. Methods Three samples of malignant melanoma tissues were ascertained by pathologic test. The total protein was extracted,separated with the method of 2D-DIGE; the difference of the proteomics was analyzed with DeCyder software; the differential-expressed protein spots were identified with MALDI-TOF/TOF-MS．Results A total of 12 protein spots were identified and categorized into 4 classes according to the function including 3 kinds of  up-regulated proteins (14-3-3 protein sigma,keratin and serpin B3)，and 1 kind of down-regulated protein (album)．Conclusion 14-3-3 protein sigma and serpin B3 are possible involved in the occurance and developmentof malignant melanoma. The mechanism of album change in the tissue of malignant melanoma remains unclear. The change of those four proteins suggests the possible pathogenesis of malignant melanoma.

Abstract:Objective To construct the vector of specific expression of the enhanced green fluorescent protein(EGFP) regulated by oxygen dependent degradation domain(ODDD) and detect the activity of ODDD in cells at limiting oxygen conditions. Methods The encoding sequences of ODDD403-601aa,ODDD549-575aa,and 2ODDD549-575aa were inserted into pEGFP-C1 respectively.Transient expression of EGFP in φA cells and relative EGFP expression were measured by RT-PCR,Western blotting and flow cytometry.Results ODDD/EGFP specific expression vector was cloned and  the function in cells was analyzed.The ODDD could specially regulate the expression of EGFP in φA cells under hypoxia conditions detected by Western blotting and the activity of ODDD was controlled by proteasome in cells.The fluorescence intensities of EGFP in φA cells under conditions of normoxia in pEGFP-ODDD401-603,pEGFP-ODDD549-575 and pEGFP-ODDD2(549-575)  groups were （6.06±1.97）％,（11.99±1.13）％,and （23.16±2.79）％,respectively,and they were lower than that of samples at limiting oxygen conditions (P<0.05). Conclusion Luciferase reporter vectors regulated by ODDD are successfully constructed.

Abstract:Objective To screen the optimal Cp G ODN for inducing the proliferation of human periodental ligament cells(PDLCs), and to provide the experimental basis for the application of Cp G ODN in treatment of periodentitis.Methods The 48 healthy premolars of young people(10-15 years old) undergoing tooth extraction for orthodontic treatment were collected.Tissue explant primary culture was used to cultivate the PDLCs in vitro.The cells from the 4th  to 5th passages were selected for experiment.1-12 Cp G ODN were respectively given to PDLCs for 24, 48, and 72 h , and the cells treated with PBS only were used as control group.MTT assay was used to detect the cell proliferation. The above experiments were repeated for 3 times.Results The human PDLCs of primary culture were obtained with tissue cultivation.The primary cultivated PDLCs showed long spindle appearance.By immunocytochemical analysis, these cells were positive to antibodies against vimentin,and negative to antibodies against ceratin, indicating themselves to be mesoderm-derived fibroblasts.The mean A values in Cp G ODN groups were all higher than those in PBS control group，and the A values of the  9th and 11th Cp G ODNs were significantly higher than that in PBS control group ( P<0.05 or P<0.01).Conclusion Cp G ODN can promote the proliferation of human PDLCs, and the 9th and 11th Cp G ODN can significantly promote the proliferation of human PDLCs.

Abstract:Objective To explore the effects of water-soluble chitosan（WSC）on angiostenosis and balloon injury of rat aorta ventralis, and to provide the basis for the development of high efficiency and low side-effect drug on prevention of angiostenosis.Methods  Seventy-five SD male rats were randomly divided into five groups: 50,100 and 200 mg/kg WSC groups, model group and  control group（n=15）.The angiostenosis animal model was established by balloon to injure rat aorta ventralis.The right common iliac artery was ligated for the rats in control group.After establishing the  animal model of angiostenosis, the WSC of 50, 100 and 200 mg/kg concentrations was respectively injected into vena caudalis of the rats in WSC groups, and normal saline were injected into vena caudalis of the rats in  model group and control group,0.4 mL every time and 1 time every day.At the 7th, 14th and 21st days after the operation, five rats in each group were randomly sacrificed, aorta ventralis were obtained and their pathological sections of HE dyeing were prepared.The pathological changes of blood vessel in each rat were observed with microscope and the ratios of lumina area to extima area at different time were counted.The inhibitory effects of WSC on angiostenosis were compared among five groups.Results Under microscope, a complete structure of aorta ventralis was observed and smooth muscle cells of tunica media were arranged in order in control group.In the aorta ventralis of the rats in model group, obvious proliferation of intima was observed at the 7th day after the operation, thickening tunica intima, thickening tunica media and localized proliferation of vessel wall were also observed at the 14th day after the operation, and many smooth muscle cells in thickening tunica intima were discovered, and smooth muscle cells of tunica media were arranged disorder and false lumen was formed at the 21st day after the operation.Compared with model group, at the 7th day after the operation, the proliferation of tunica intima was obviously lessen in experiment group, and especially in 100 and 200 mg/kg WSC groups.At the 14th day after the operation, thickening tunica intima and media were not obviously observed, but in  200 mg/kg WSC group, large smooth muscle cells with disorder arrangement werestill visible in tunica media.At the 21st day after operation, the vessel wall of aorta ventralis was normal in all WSC groups, especially in 200 mg/kg WSC group.The comparison of the relative lumen area showed that the variation of the relative lumen area of the rats in control group did not change significantly following the time after operation (P>0.05); but compared with control group, the lumen area variation of the rats in model group at the 7th, 14th and 21st days had very significant difference (P<0.01).50 mg/kg WSC could enlarge the lumen area of the rats at the 14th and 21st days, but compared with control group, the difference was still very significant (P<0.01).100 mg/kg WSC could enlarge the lumen area better than 50 mg/kg WSC did; and at the 14th and 21st days, the difference between WSC groups and control group was still significant (P<0.05).200 mg/kg WSC was the best concentration on the enlargement of the lumen area after 14 d, and there was no significant difference compared with control group (P>0.05).Compared between the different days of all groups, there were no significant differences（P>0.05）.Conclusion WSC could significantly inhibit the angiostenosis after balloon injury of rat aorta ventralis and the inhibitory effect could be the best at the concentration of 200 mg/kg and more than 14 d.

Abstract:Objective To study the effects of long-term exposure to formaldehyde on cognition and motor function of mice,and explore the potential mechanisms.Methods  48 Kunming mice were randomly divided into 4 groups: negative control and low, middle and high dose formaldehyde groups(21.0, 42.0 and 84.0 mg/m3).The mice were exposedto formaldehyde by inspiration in a static total enclosure chamber for 12 weeks. Before exposed to formaldehyde and at 4th week, 8th week and 12th week, cognition and motor function of mice were determined.At 12th week, the mice were killed, the activities of succinate dehydrogenase(SDH),Na+-K+ ATPase and Ca2+-Mg2+ATPase in the brain tissues were detected.Results   At the test of initiative escape, the times of initiative escape in high dose group were significantly decreased compared with control group at the 12th week（P<0.05).At the test of passive escape,at the 12th week, the time of passive escape of the mice in middle dose group and high dose group was increased, and there was significant difference compared with control group（P<0.05 or P<0.01).With extending of exposure to formaldehyde, the time of passive escape had changed.At 12th week, the escaping time in high dose group was significantly increased compared with the 4th week and the 8th week（P<0.05).The   SDHactivities were decreased in form aldehyde groups, and there was significant differences between high dose group  and control group (P<0.05); the activities of Na+-K+ATPase and Ca2+-Mg2+ATPase were decreased in formaldehyde groups, but there were no significant differences compared with control group (P＞0.05).Conclusion Long-term exposure to formaldehyde can  decrease of cognition and motor function and the change of energy metabolism of brain tissue  of mice,indicating that the decrease of energy metabolism may be the potential mechanisms of decrease of cognition and motor function.

Abstract:Objective  To investigate the effect of the expression of sodium ferulate(SF) on the related protein of Rho/Rock signal pathway and to clarify the protective effects on liver tissue and myocardium tissue.Methods Twenty male Wistar rats were divided into normal control group, model group, positive medicine group,and sodium ferulic group.Besides normal control group,the rats in the  other groups were injected with 15 mg/kg Iso to induce rat myocardial fibrosis.2 d later, except control and model groups, the rats in the other groups were given SF and Bena zepril.The  change of collagen content in myocardium was analyzed by Masson staining.The expressions of Rho A and Rock  mRNA were analyzed by RT-PCR.The expression of Rock  protein was analyzed by immunohistochemistry. The Raman spectrum was used to detect the liver tissue injury.Results Compared with control group,the expressions of RhoA and Rock mRNA were increased after injection with Iso for 3 weeks(P<0.01);compared with model group,the expressions of Rho A and Rock mRNA and protein were decreased after injection with SF (P<0.01).The Raman spectrum results showed that the peak value in SF group shifted to the right with one unit compared with model group,and tended to normal control group.Conclusion SF can not only protect the heart against fibrosis by inhibiting Rho/Rock pathway, but also can protect liver damage.

Abstract:Objective To establish a new method to determine the contents of  matrine and oxymatrine in Qingre Tonglin Tablet by high performance liquid chromatography(HPLC),in order to optimize the quality evaluation and control of Qingre Tonglin Tablet. Methods HPLC method was adopted.Matrine and oxymatrine standards were the control substances and Qingre Tonglin Tablets were the experiment samples.Using Agilent 1200 high performance liquid chromatograph,the determination was performed on Genimi-C18 (5  μm,4.6 mm×250 mm) column with mobile phase consisting of phosphate buffer solution (PH3.0) - methanol (93∶7) at the flow rate of  0.8 mL/min.The column temperature was 35℃.The detection wavelength was 220 nm and the injection volume was 10  μL.For the methodological study,thelinearity,specificity,stability,precision and repeatability and accuracy of the HPLC method were tested. Results The regression equation of matrine was Y=1102.1X+15.423,showing good linearity (r=0.999 9 ) in the range of 0.311　8-4.677　0 μg.the average recovery rate was 97.1% (RSD = 0.24%,n=6). The regression equation of oxymatrine was Y=1106.6X+5.040 2,showing good linearity (r=0.999　6) in the range of 0.029　8-0.446 4 μg; the average recovery rate was 98.4% (RSD=1.52%,n=6). In the tests of stability,precision and repeatability,the RSD of matrine and oxymatrine were respectively 1.21%,1.47%; 0.13%,2.76%; 0.48% (RSD of the sum of matrine and oxymatrine). Conclusion HPLC method issimple,sensitive, reliable,specific and has satisfactory separation results,and it is competent for the quality control of QingreTonglin Tablet.

Abstract:Objective To investigate the antibacterial effect of room curing polymethylmethacrylate(PMMA) material adding nano-silver base inorganic antibacterial agent and to detect the changes of its mechanical property.
Methods Nano-silver base inorganic antibacterial agent was added to the room curing PMMA material in the range of 0.5%-3.0% at an interval of 0.5% by ball milling to make specime.Antibacterial rates of the specimes were detected by film method.Bending strength,impact strength,and wear resistance of the specimes were respectively detected on electronic universal testing machine,impact test machine and friction and wear test machine.Results The antibacterial rates of Streptococcus mutans and Candida albicans were more than 50% when antibiotics content was 1.0%.The antibacterialrates of Streptococcus mutans and Candida albicans were more than 90% when the antibiotics content was 2.5%.The three mechanical properties were increased compared with control group when the antibacterial agents were in the range of 1.0%-1.5%.Then the three mechanical properties were decreased with the increasing of antimicrobial concentration.When the antibiotics content was 2.0%,the wear resistance had significant difference compared with  control group  (P<0.05); when the antibiotics content was 2.5%,the bending strength and impact strength had significant difference  compared with  control group(P<0.05).Conclusion The antibacterial effect of room curing PMMA adding nano-silver base inorganic antibacterial agent is ideal.The antibacterial rate is increased gradually with the increasing content of antibacterial agents.There is no significant effect on the mechanical properties of room curing PMMA material,but the antibacterial  effects are satisfied when the content of antibacterial agents is 2.0%.

Abstract:Objective To discuss the relationship between the expression of death-inducing signaling complex (DISC) and apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL) in malignant glioma cells,and to explore the mechanism of resistance of TRAIL initiated apoptosis. Methods The primary cultured cells were isolated from human malignant glioma tissues,and  the level of apoptotsis was detected by acid phosphatase assay after the  cells  treated with different doses of TRAIL; and  the level of DISC protein was determined by Western blotting.Results The primary malignant glioma cells GC417,GC321 and GC125 were isolated and cultivated,and the sensitivities of the cells to TRAIL were different,GC321 (0.12±0.01  vs  0.51±0.02) and GC125 (0.22±0.01  vs  0.36±0.01) were sensitive to TRAIL,there were significant differences compared with control group(P<0.01);but GC417 (0.24±0.01  vs  0.23±0.02 ) was resistant to TRAIL.The results of Western blotting showed that the expressions of DISC proteins were various,and the expressions of GC321 and GC125 were increased,and the expression of GC417 was decreased.Conclusion The responses are different according to the different primary malignant glioma cells to TRAIL-induced apoptosis,and the expressions of DISC proteins are either different; there is relationship between the decreasing of the  expressions of DISC proteins and the  resistance of TRAIL initiated apoptosis.

Abstract:Objective  To study the neuroprotective effects of Humanin(HN) on exciatory neurotoxicity  based on inhibiting  the expressions of NMDA receptor NR1 subunits and  phosphorylation of NR2B subunits, and to explore the possible mechanisms.Methods Primary cultured  cerebral cortical neurons of Wistar rats were used to set up excitatory neuotoxicity model and  were divided into control group, NMDA group,NMDA+MK-801 (MK-801) group and NMDA+HN group.The expressions  of NR1 subunit proteins of NMDA receptors  were detected by flow cytometry  (FCM).The levels of NR2B subunits of NMDA receptors phosphorylation at Tyr1472 in various  groups were detected by Western blotting.Results  The results of inverted phase contrast microscope showed that in NMDA group the cerebral cortical neurons were obvious neuronal loss,few cells became round and there was halofuzzy around the cells and the neurices were mostly gone, and the cells showed obvirous  excitatory neurotoxicity.The mean fluorescence intensity value of NR1 subuint protein of NMDA receptor  dsignificantly was increased  compared with control group（P<0.05）,the mean fluorescence intesity values of NR1 subunit protein were  reduced in MK-801  group and HN group compared with NMDA group（P<0.05）.The Western blotting results showed that there were no significant changes of total NR2B subunit proteins in various groups.The level of NR2B phosphorylation at Tyr1472 induced by NMDA  was increased significantly compared with control group（P<0.05）, while the pretreatment of HN(1  μmol/L) attenuated the level of phosphorylated of NR2B subunit  in primary cultured rat cerebral cortical neurons compared with control group（P<0.05）.Conclusion The up-regulation of the expression level of NR1 subuint protein of NMDA receptor and the  increaing  of the level of the phosphorylated of NR2B subunit  might be responsible for the excitatory       neurotoxicity induced by NNDA.HN might contribute to the neuroprotection against excitatory neurotoxicity  by inhibiting the expression of NR1 subunits and reduction of phosphorylated of NR2B.

Objective To probe the influence  of erythropoietin (EPO) on ischemic size and infarction size of  rats with  myocardial ischemia-reperfusion (I/R) injury   and to clarify the mechanism of the protective effect of EOP on myocardial I/R injury.Methods 24 healthy male SD rats were randomly divided into sham-operated group(n=8), I/R group(n=8) and EPO group(n=8).In sham-operated group, the 6-0 thread was passed through the left anterior descending(LAD) coronary artery of rats  without further procedures; in I/R group, the LAD coronary artery were ligated and embedded slack line in the ligature line and reperfusion was performed 45 min later; in EPO group, the rats  had the same surgical procedures as I/R group and were administered by intraperitoneal injection of recombinant human erythropoietin(rhEPO) 5 000 U/kg.The hemodynamic parameters including systolic  pressure(SP), diastolic  pressure(DP),left ventricular systolic pressure(LVSP),left ventricular diastolic pressure(LVDP),left ventricular end-diastolic pressure(LVEDP),maximal rate of increase of ventricular pressure(dp/dtmax ) amd maximal rate of decrease of ventricular pressure  (dp/dtmin)were determined by multiple channel electrophysiolograph after 24 h reperfusion.The apoptotic index of cardiomyocytes was investigated by TdT-mediated dUTP nick end labeling(TUNEL) method.The ischemia and infarction size of hearts were measured by double staining with TTC-Evan’s blue dye.Results After 45 min ischemia and 24 h reperfusion, there were no significant differences in SP,  DP and LVSP between three groups(P>0.05). Compared with sham-operated group, LVDP and LVEDP were significantly increased while dp/dtmax and dp/dtmin were reduced remarkably in I/R group(P<0.05);  compared with I/R group, LVDP and LVEDP were decreased significantly but dp/dtmax and dp/dtmin were increased remarkably in EPO group(P<0.05).The results of the myocardial TUNEL staining revealed that the TUNEL positive cardiomyocytes were increased dramatically after I/R injury;but compared with I/R group, the apoptosis was reduced significantly in EPO group (P<0.05).The results of the myocardium staining showed that the myocardial infarction size was reduced in EPO group compared with I/R group (P<0.05).Conclusion EPO can significantly protect the heart from I/R injury through improving the hemodynamic status and heart function following the reduction of  cardiomyocyte apoptotic and ischemic size as well as infarction size of the left ventricle of the  rats with I/R injury.

Abstract:Objective  To explore the effects of cyclosporine A(CsA) and porphyromonas gingivalis-lipopolysaccharide(Pg-LPS) on the area of buccal gingiva of the firstmolar of mandibular and CEJ-ABC distance of rats.Methods Forty rats were equally divided into eight groups.There were 5 rats in each group.The first molar ofmandibular of rats in various groups were injected,from the buccal side,with saline,CsA（2.0 mg/kg）,LPS（10.0,1.0,0.1  μg）,CsA+ LPS（10.0  μg）,CsA+ LPS（1.0  μg）,CsA+ LPS（0.1  μg）,respectively.After 30 d,the mandibles were then  dissected and sectioned for histological and immunocytochemical analysis by haematoxylin andeosin staining,the changes of the distance of CEJ-ABC and the area of buccal gingiva in various groups were observed.Results CsA（2.0 mg/kg） increased the area of thebuccal gum of first molar and the distance of CEJ-ABC，but there was no statistically significant difference compared with control group（P>0.05）.LPS （10.0  μg）alone or combined with CsA  could damage the alveolar bone and increase the distance of CEJ-ABC，and there were  statistically significant differences compared with control group（P<0.05）;but  there was  no significant difference when treated with LPS (1.0,0.1  μg) alone or combined with CsA;and  no obvious change in gingival was found following all of the treatments（P>0.05）.Conclusion  Low dose of CsA（2.0 mg/kg） has no damage on the periodontal tissues of rats,and such a level of CsA may not decrease the periodontal breakdown induced by  high dose  of LPS(10.0 μg).

Abstract:Objective To evaluate the inhibitory effect of Flavones of Buckwheat Compound(FBC )on the histomorphologic changes of the kidney tissue in type 2 diabeticrats,and to explore the mechanism of FBC on diabetic nephropathy(DN).Methods The type 2 diabetic male SD rat model was induced by feeding high-fat diet and intraperitoneal injection with streptozotocin (STZ).The rats were randomly divided into five groups: model group(distilled water,10 mL/kg/d),positive medicine group(XKW,0.83 g/kg/d),low and high doses FBC groups(FBC,0.6 and 1.2 g/kg/d),and normal control group(distilled water, 10  mL/kg/d).The rats in each group  were gavaged daily for eight weeks.At the end of the experiment,the level of fasting blood glucose(FBG) was detected and the index of kidney was observed.HE and Masson staining were used to observe the morphological changes of kidney tissue under optical microscope.The changes of the kidney tissue ultrastucture were observed using   electron microscope.Results  Compared with model group,the level of FBG and kidney index in FBC groups were significantly decreased（P<0.01).However there were no obvious differences in these indexes between control group and FBC groups.And FBC could inhibit the increasing of mesangial matrix and the thickening of basement membrane in kidney of diabetic rats,and relieve glomerular sclerosis and renal tubular epithelial cell vacuolation.Conclusion FBC could inhibit the increasing of mesangial matrix and basement membrane thickening of diabetic rats.The mechanism is related to inhibiting mesangial cell proliferation and glomerular basement membrane formation and improving the increasing of diabetic glomerular volume and hardening.

Abstract:Objective  To observe the effects of Soft Capsule of Compound Oil of Jujube,Arboruitae,and Gardenia (COJAG) on serum estrogen hormone,blood rheology,blood fats,as well as body weight and viscera indexes in castrated female rats. Methods 45 bilateral oophorectomy female rats were divided into COJAG high,middle and low doses groups,estrogen group,and model group; other 10 rats were used as  sham operation group. The blood fats,blood rheology,body weight,viscera indexes,serum estrodiol,follicle-stimulating hormone and luteinizing hormone were determined. Results Compared with model group, the serum E2 was significantly increased (P<0.01),FSH (P<0.05) and LH (P<0.01) were reduced; TG,TC,LDL,and C/HDL-C were significantly reduced (P<0.05),HDL-C was increased (P<0.05); the body weight of the female castrated rats was significantly reduced and their thymus,pituitary,uterus,adrenal and other organs’ viscera indexes were increased (P<0.05 or P<0.01). The ηb and ηp in high,middle and low shear rates of whole blood viscosity,plasma viscosity were significantly lowered (P<0.05),ESR was speeded up (P<0.05),and the Hct and Fib were lowered (P<0.05) in the female castrated rats in COJAG groups. Conclusion COJAG can decrease blood fat,improve blood rheology,reduce weight,and increase viscera index; these roles are probably related to the increase of estrogen content in blood and(or) its rich polyunsaturated fatty acids.

Abstract:Objective To optimize the prescription and preparation of recombinant human acidic fibroblast growth factor(rhaFGF) liposomes,and to study the physical and chemical properties. Methods The encapsulation efficiency and biological activity of rhaFGF liposomes were evaluated by single factor test. Freeze-drying-rehydration was used to obtain the optimum prescription and preparation of rhaFGF liposomes. Results The encapsulation efficiency of rhaFGF liposomes was (81.79±2.51)%,the physical stability index (KE) was (1.050±0.342)% and the bioactivity was (6.521 0±0.617 1)×105U/mL. The particle size was about 100 nm uniformly. Conclusion The preparation of rhaFGF liposomes is optimized,and these liposomes have high encapsulation efficiency,good physical stability and high bioactivity at 4℃.

Abstract:Objective  To study the effect of Lipid-lowering Mixture(LM) on blood lipid profile and blood hemorheology of hyperlipidemia rats，and to elucidate the mechanism of LM on regulating blood lipid and hemorhelogy. Methods  60 male Wistar rats were randomly divided into  normal control group, model control group,simvastatin group and LM high,middle, low doses groups,and there were 10 rats in each group.The rats in normal control group were fed with normal  diet,and  the rats in the other groups were fed with a high fat diet to establish hyperlipidemia rat models, and during the modeling the drugs were  used for prevention.2 mL physiological saline were given to the rats in control group and model group,and the rats in simvastatin group  were given 7.2×10-4g/mLsimvastatin suspension solution 2 mL, and the rats in LM low, middle,high doses groups were given 0.25,0.50,1.00 g/mL decoction 2 mL,the once  daily dose  lasted for 10 weeks.After 10 weeks,the rats in 6 groups were  eyeballed blood, the blood lipid profile and blood hemorheology of each  animal in various groups were measured by full automatic biochemical analyzer and full automatic blood analyzer,respectively.Results Compared with model group, after  treatment, the levels of the cholesterol (TC ), triglyceride ( TG ),low density lipoprotein ( LDL-C ) and  apolipoprotein B ( ApoB ) in LM  groups were reduced; the high density lipoprotein ( HDL-C ), apolipoprotein A ( ApoA ) levels and apoA / apoB ratio were increased, the differences were statistically significant ( P<0.05 or P<0.01).Compared with model group, the plasma viscosity（ηp）, whole blood viscosity ( low, medium, high cut ) （ηb）, index of erythrocyte aggregation ( IEA ), hematocrit (HCT ),and erythrocyte electrophoresis time ( EET ) in LM groups were decreased, the index of erythrocyte deformability ( IED ) was increased, the differences were statistically significant ( P<0.05 or P<0.01).Conclusion LM can modulate blood lipid metabolism and blood viscosity, poly, and prothrombotic state of the experimental hyperlipidemia rats, which has better regulation and inhibition  on blood lipid and blood flow abnormalities.

Abstract:Objective  To optimize the fermentation condition of large scale fermentation of human oncostatin M(hOSM)  with engineering strain of Pichia pastoris. Methods The optimal pH condition of hOSM expressed in the methylotrophic yeast Pichia pastoris X-33 in tube-scale was determined and fermentation of hOSM was performed in an 80 L fermentor in a fed-batch mode for three times.The pH value, culture medium, dissolved oxygen, temperature, methanol feeding speed, initial biomass and etc were focused mainly.hOSM was analyzed by SDS-PAGE.Results  In the FM21 medium of pH 5.0, when a cell yield of 190 g/L wet weight was achieved, methanol induction phase began.In the fermentation broth of pH 5.5, the temperature was controlled at 28 ℃, a stirring speed of up to 650 r/min, the pressure of fermentor was 10 P, dissolved oxygen above 25% and the supply speed of methanol was 9.3 mL?h-1?L-1 initial fermentation volume, after methanol induction for about 36 h, the expression level of hOSM reached 280 mg/L. Conclusion  OSM is successfully obtained through genetic engineering for large scale production of hOSM for following biological functions.

Abstract:Objective  To compare the effects of different doses of sodium fluoride(NaF) and hydrogen peroxide(H₂O₂) on expression of vascular endothelial growth factor(VEGF) in fibroblasts of rats,and to explore a simple and effective way to promote the expression of VEGF,and in order to provide some theoretical reference to optimize the dermis culture condition in tissue engineering.  Methods  The rat fibroblasts were isolated by enzyme digestion,and they were cultivated respectively with DMEM medium contained different doses of sodium fluoride （0.05 and 0.50 mmol/L ）and H₂O₂（10  nmol/L1and 10 μmol/L）,or with both two factors（NaF 0.05 mmol/L,H₂O₂ 10 μmol/L）.The proliferation ability was evaluated by CCK-8 assay at 0,24 and 48 h after  the fibroblasts were treated with these factors.And the effects of  NaF and H2O2 on the expression of VEGF were evaluated by ELISA assay and immunohistochemical method at 48 h.Results  There was no evidently inhibitory effects  of  different doses of NaF and H₂O₂used alone or combined   on the proliferation of fibroblas,there was  no significant difference compared with  blank control(P>0.05).The expressions of VEGF in both NaF group and H2O2 group were greatly improved,there was significant difference compared with   blank control group(P<0.05).However,the expression of VEGF in the fibroblasts treated  with  combined two factors was most enhanced,which could increase 40%(P<0.05). Conclusion The combination of NaF and H2O2 can keep the  normal proliferation  activity of fibroblasts,and promote the expression of VEGF effectively.

Abstract:Objective  To establish human squamous carcinoma cell line Hep-2 stably overexpressing exogenous ANO1，and to  observe its effects on the biological characters of Hep-2 cells.Methods Primers were designed.DNA was extracted from clinical specimens of patients with head and neck squamous cell carcinoma, and taken as template for PCR.Gene sequences of ANO1 fragment were gotten.ANO1 fragment was inserted into the pSG-5 plasmid, which was transfected into Hep-2 cells.These Hep-2 cell lines with ANO1 stable overexpression were regarded as experiment group,and those Hep-2 cell lines transfected with blank plasmid were regarded as control group.The expression of ANO1 was detected by Western blotting and immunofluorescence assay in the Hep-2 cells after transfection.Boyden chamber invasion assay and adhesion assay were used to observe the effects of ANO1 overexpression on the biological characters of Hep-2 cells.Results The PSG-5 plasmid containing ANO1 fragments was successfully constructed.The expression of ANO1 in Hep-2 cell line after transfection was observed by Western blotting.The result of Western blotting showed that the target band occurred in each group, and 100 000 band was darker in experiment group.Indirect immunofluorescence assay showed that the bright green fluorescence presented in Hep-2 cells with stable overexpression of ANO1; while darker, lighter green fluorescence in Hep-2 cells after transfection with the blank plasmid.The percentage of ANO1 overexpressing Hep-2 cells migrating through the membrane and percentage of ANO1 overexpressing Hep-2 cells adhesion of cells were greater than those in  control group（P<0.05）.Conclusion  The Hep-2 cell line with ANO1 stable overexpression is successfully established.Stable ANO1 overexpression can improve the migration of Hep-2 cells.

Abstract:Objective  To observe the neuroprotective effect of Neanthes japonica (Izuka) fibrinolytic enzyme (NJF) on rat models of focal cerebral ischemia and to explore its mechanism.Methods  60 Wistar rats were randomly divided into sham　operated group, model group, NJF treatment groups (0.25, 0.50 and 1.00 mg/kg, respectively) and Urokinase group (UK, 15 000 U/kg ).The rat model of middle cerebral artery occlusion (MCAO) was produced by intraluminal suture method. The neurological deficits score was measured 24 h after the operation, the cerebral infarction was detected by TTC staining, the brain water content was determined by wet and dry weights, and the malondialdehyde (MDA) levels and superoxide dismutase (SOD) activities were measured by chemical colorimetric assay.Results  Compared with model group, the neurological deficits scores of rats in all NJF groups were significantly decreased (P<0.05), the cerebral infarction area was significantly reduced (P<0.001), the brain water contents were decreased (P<0.05 or P<0.01), the MDA levels were decreased (P<0.001) and the SOD activities were increased (P<0.05 or P<0.01) significantly in hippocampus of the cerebral ischemia rats.There were no significant differences in the cerebral infarction area and the brain water content of rats between   0.50 mg/kg  NJF group and   15 000 U/kg UK group.Conclusion  NJF posseses neuroprotection for MCAO and reperfusion model rat.The neuroprotection of NJF may be related to inhibiting lipid peroxidation and increasing the activities of endogenous antioxidant defense enzymes.

Abstract:Objective To study the effects of puffball on proliferation and collagen synthesis on rat skin fibroblasts, and to investigate the  mechanism of  enhancing wound healing  of  active constituent of puffball     at the cells level.Methods The rat skin fibroblasts were divided into 5 groups, all the cells were treated with puffball(37.5,75.0,150.0 and 300.0 mg/L), except for control group,for 24,48 and 72 h,respectively.And then MTT was used to detect the proliferation, and the content of collagen were measured by double antibody sandwich ELISA.Results  Compared with control group, the cells treated with different concentrations (37.5,75.0,150.0 and300.0 mg/L ) of puffball showed a significant rise both in its proliferation and the ability of collagen synthesis (P<0.01).Furthermore, in time-dependant manner,  the proliferation rate of cells and collagen synthesis ability  wer  significantly increased after treated for 24,48,72 h,especially in 72 h(P<0.01). Conclusion  Puffball with different doses  can enhance the proliferation and collagen synthesis in  skin fibroblasts of rats.

Objective  Below the national health standards,to observe the changes of the blood cell count and P16 /CDKN2A protein content in plasma of workers who long-term exposed to benzene series,and to provide theoretical basis for better monitoring and supervision in benzene series pollution.Methods 50 workers which worked on operating environment more than 2 years were selected as exposed group in one printing plant in Shenzhen,and  50 administrative and logistic staffs in the same printing plant were used as control group.The inspections were taken on white blood cell count,red blood cell count,hemoglobin content and platelet count in
 two groups by blood routine examination.The plasma P16 /CDKN2A protein content in two groups were detected by ELISA assay.The concentrations of benzene,toluene and xylene in exposed group in the operating and non-operating environment were determined at the same time.Results The benzene,toluene and xylene levels in exposed and control groups were lower than the national health standards.The white blood cell count,red blood cell counts,hemoglobin contents and platelet counts in two groups were all in normal range and had no significant differences between two groups(P>0.05).The average content of plasma P16 /CDKN2A protein of the workers in the exposed group was lower than that in control group,but there was no significant difference between two groups(P>0.05).Conclusion  In the exposure condition,long-term exposure to low concentrations of benzene series has no significant effects on blood cell count and plasma P16 /CDKN2A protein content of the workers.

Objective To explore the association  between APOL3 and APOL1 gene polymorphisms in APOL gene family and schizophrenia,and to lay a foundation for etiology research on schizophrenia.Methods  97 families composed
 with schizophrenia patients who were Han nationality and their healthy parents in  Jilin Province from China were chosen.The single nucleotide polymorphism (SNPs) of rs132630 and rs2239785 sites of APOL3 and APOL1 gene were detected by  polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP)  assay.The allelic and genotypic frequencies fit Hardy - Weinberg Law were detected by  goodness of fit χ2 test analysis; the relative risk  of the haplotypes(HRR)  and transmission disequilibrium test(TDT) were detected by UNPHASED software.Results The distribution of genotypic frequencies of rs132630 and rs2239785 did not deviate from Hardy-Weinberg equilibrium  in both patient and parent groups ( P>0.05).The  HRR  test showed there were no significant differences of allelic frequencies  of rs132630 and rs2239785 between patient  and parent groups (P>0.05).The  TDT  showed the alleles of rs132630 and rs2239785 transmitted from heterozygotic parents to affected offsprings did not bias from 50% ( P>0.05).Conclusion The polymorphism of rs132630 at APOL3 gene and rs2239785 at APOL1 gene may not be associated with schizophrenia.
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Objective To study the expressions of galactose Galectin-3 and integrin ανβ3 mRNA in glioma cells  and their roles in the occurrence and the malignancy progress  of glioma.Methods  In situ hybridization was used to detect the expressions of Galectin-3 and integrin ανβ3 mRNA in 5 normal brain tissues and 40 patients with different grades gliomas , and the relationship between them was analyed. Results The expressions of Galectin-3 and integrin ανβ3 mRNA in normal brain tissue were negative.Galectin-3 mRNA mainly expressed in the cytoplasm and membrane of tumor cells.In 23 cases ofⅠ, Ⅱ grade glioma specimens, 2 cases were positive,4 cases were weakly positive and 17 cases were negative, the positive rate was 29.09%(6/23), LI value was (4.80±3.22)%;in  17 cases of Ⅲ, Ⅳ grade glioma specimens,4 cases were strongly positive,6 cases were positive ，1 case was weakly positive and 6 cases were negative，the positive rate was 64.71%(11/17), LI value was (27.30±22.72)%; there was significant difference in the positive rate betweenⅠ，Ⅱ grade and Ⅲ, Ⅳgrade glioma(P<0.05). The integrin ανβ3 mRNA mainly expressed in the cytoplasm and membrane of tumor cells. In 
23 cases of Ⅰ, Ⅱ grade glioma specimens 3 cases were positive expressin of integrin αυβ3,4 cases were weakly positive, 16 cases were negative, the positive rate was 34.43% (7 / 23), LI value was (5.35 ± 3.79)%; in  17 cases of Ⅲ, Ⅳ grade glioma specimens,4 cases were strongly positive, 6 cases were positive, 3 cases were weakly positive, and 4 cases were negative，the positive rate was 76.47% (13/17), and LI value was (29.24± 21.10)%; there was significant difference in the positive rate betweenⅠ，Ⅱ grade and Ⅲ, Ⅳgrade glioma(P<0.05).The linear correlation analysis showed that the expression rates of Galectin-3 and integrin ανβ3 mRNA  were positively correlated (P<0.05).Conclusion The expressions of Galectin-3 and integrin ανβ3 mRNA  in Ⅰ, Ⅱ grade gliomas are significantly lower than those of Ⅲ, Ⅳ grade glioma,which indicates that the detection of  Galectin-3 and integrin  ανβ3 mRNA  can be used as an indicator to judge malignant biological behavior of human brain glioma at molecular level.

Objective  To study the different positive end-expiratory pressure (PEEP) on pulmonary shunt fraction and oxygenation during one-lung ventilation (OLV), and to explore the best PEEP value in OLV.Methods  Thirtypatients with ASA Ⅰ-Ⅱ level elective thoracotomy with OLV were selected.The blood gas analysis were performed
 in the lateral position at ventilation 30 min (T1), OLV 30 min (T2), 5 cmH₂O OLV PEEP 30 min (T3), and 10 cm

H₂O OLV PEEP 30 min (T4).At each time point, the PaO₂, PaCO₂, SaO₂, pH, HR, mean arterial pressure (MAP), and peak airway pressure (Ppeak) were recorded, and the intrapulmonary shunt(Qs / Qt) was calculated.Results Compared with two-lung ventilation, the PaO₂ was decreased and the Qs / Qt was increased significantly (P<0.05), the PaO₂ was increased from (200.20 ± 145.25) mmHg to (299.55±138.83) mmHg (P<0.05) and the Qs / Qt was decreased (P<0.05) at the duration of T2 to T3; the PaO₂ was decreased from (299.55 ± 138.83) mmHg to（237.30 ± 135.57） mmHg(P<0.05) and the Qs / Qt was increased (P<0.05) at the duration of T3 to T4; the Ppeak value was increased from T2(21.15±3.60) cmH2O to T4(27.20±3.78) cm

H₂O (P<0.05)  in OLV.Conclusion During OLV, 5 cmH2O PEEP applied in ventilated lung can increase the

PaO₂, decrease the Qs / Qt and prevent the occurrence of hypoxemia.It is more appropriate PEEP value.

Objective  To detect  the expression of endothelial derived gene-1 (EG-1) gene  in thyroid papillary carcinoma tissue by real-time quantitative PCR and to investigate  the relationship between EG-1 and the ocurrence and  development of thyroid papillary carcinoma. Methods Real-time quantitative PCR was used to detect
the expressions of EG-1 gene in 38 thyroid papillary carcinoma patients ( case group) and 15 thyroid adenoma patients (control group),and the expression level of EG-1 was detected by immunohistochemisty.
Results The expressions of EG-1 mRNA and protein in thyroid papillary carcinoma tissue  were significantly higher than those in adenoma  tissue（P<0.05）.The expression of EG-1 mRNA had significant relationship with the pathological degree in thyroid cancer(P<0.05), but there was no correlation with the age and the gender of the patients(P>0.05).Conclusion The expression of EG-1 gene in thyroid papillary carcinoma tissue is obviously higher,EG-1 gene  may be important in the occurrence and  development of thyroid cancer.
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Objective  To investigate the correlation between 4 single nucleotide polymorphisms（SNPs） of vitamin D receptor (VDR) gene and type 2 diabetes mellitus （T2DM） in Ningxia Han population and to provide theoretical basis for  T2DM prevention.Methods  Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method was carried out to examine the SNPs at rs1544410 (G>A)，rs757343 (G>A)，rs731236 (T>C) and rs739837 (G>T) sites of VDR gene  in 201 T2DM patients (T2DM group) and 210 healthy individuals (control group) in Ningxia Han population.The χ2 test was used to compare the allelic frequencies and genotypic frequencies between two groups.The SHEsis online haplotype analysis software was applied.
Results The genotypic frequencies of rs1544410 and rs757343 had significant differences between T2DM group and control group (P<0.05).The frequency of allele G was higher in T2DM group (93.5%) than that in control group (89.3%), and the difference between  two groups had statistically significant difference (P<0.05).The OR
［95%CI］ value was 1.738 ［1.055-2.865］.There were 16 haplotyes in 4 SNPs in all and  the frequency of haplotype GGCT had significant difference between T2DM group (10.0%) and control group (6.1%)(P<0.05),
with OR ［95%CI］ = 1.723 ［1.03-2.883］.Conclusion The allele G of rs1544410 and the haplotype GGCT of VDR gene may be associated with T2DM susceptibility in Ningxia Han population.
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Objective To detect the expressions of MMP-14 and TIMP-2 in colorectal carcinoma tissues, and investigate the relationship between MMP-14, TIMP-2 and clinicopathological features.Methods  60 paraffin-embedded cancer specimens   from operated patients with colorectal carcinoma   were selected, and 20 normal mucous memb
ranes of colon and rectum were selected as control.The expressions of MMP-14 and TIMP-2 were detected by S-P immunohistochemistry.The relationship between the expressions of MMP-14, TIMP-2 and clinicopathological parameters were analyzed.Results  The expression rates of MMP-14 and TIMP-2 were 86.7% and 80.0%, respectively, in colorectal carcinoma tissues and were significantly higher than those in control group (P<0.01).The expression of MMP-14 was closely correlated with the depth of tumor invasion, lymph node metastasis and Dukes stage.The expression rates of MMP-14 in serosa group, lymph node metastasis group and Dukes C＋D stage were 89.5%, 94.1%, and 94.3%, respectively, and were significantly higher than those
 in non-serosa group, no lymph node metastasis group and Dukes A＋B stage (68.2%, 73.1%, and 76.0%)（P<0.05).The expression of TIMP-2 was closely correlated with the depth of tumor invasion, lymph node metastasis and Dukes stage.The expression rates of TIMP-2 in serosa group, lymph node metastasis group and Dukes C＋D stage were 55.3%, 64.7%, and 62.9%, respectively, and were significantly lower than
those in non-serosa group, no lymph node metastasis group and Dukes A＋B stage (81.8%, 88.5%, and 92.0%)(P<0.05).There was negative correlation between the expressions of MMP-14 and TIMP-2 in colorectal carcinoma tissues(rs＝－1.0, P<0.05). Conclusion  The high expression of MMP-14 in colorectal carcinoma tissues may contribute to tumor invasion and metastasis and  TIMP-2 may play an inhibitory effect  in the development of colorectal carcinoma. The imbanlance of MMP-14 and TIMP- 2 may be one of the mechanisms of tumor invasion and metastasis.
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Objective To investigate the coronary artery calcium scores of subjects with different 25(OH)D levels and to clarify the relationship between 25(OH)D level and coronary artery calcification.Methods A total of 184 subjects were
evaluated by multi-slice spiral CT coronary artery calcium imaging and 25(OH)D levels testing.The subjects were divided into two groups according to 25(OH)D level: 25(OH)D deficient group(＜15 μg?L-1) and 25(OH)D  normal  group≥(15 μg?L-1). The average coronary artery calcium scores of the two groups were compared and the relationship between coronary artery calcium scores and 25(OH)D levels of all 184 subjects was analyzed.
Results There were 120 subjects in normal group, whose average coronary artery calcium score was 43.9±91.7, and 55 subjects out of  the group (45.8%) scored zero.The 25(OH)D deficient group had 64 subjects, whose
 average coronary artery calcium score was 200.0±113.5, and 21 out of the group (32.8%) scored zero.The average coronary artery calcium score in normal group was much lower than that in deficient group, and the number of zero-score subjects in normal group was also more than that in deficient group; the differences between two groups were statistically significant(P<0.05).There was negative relationship between coronary artery calcium scores and 25(OH)D levels (r=－0.147, P<0.05).Conclusion The 25(OH)D level is negatively correl
ated to coronary artery calcium score.The deficiency of 25(OH)D  can cause coronary artery calcification.
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Abstract:Objective To compare the therapeutic efficacy and adverse effects of melphalan and prednisone plus thalidomide regimen ( MPT ) with melphalan and prednisone regimen ( MP ) in the treatment of patients with multiple myeloma ( MM ).Methods 37 elderly patients  newly diagnosed as MM  were treated with MPT or MP.The patients were divided into two groups according to the treatment regimen.MPT group (20 patients):  melphalan 4 mg.m-2?d-1, d 1-7
, prednisone 40 mg?m-2?d-1, d 1-7,thalidomide 100-200 mg?d-1,every 28 d for 1 cycle.MP group (17 patients): the doses of melphalan and prednisone were the same as that in MPT group.The efficacy was evaluated after 4-6 cycles of treatment.Results The overall response rate (ORR ) of the patients in MPT group was significantly higher than that in MP group (65% vs 41.2%，P<0.05). The median time to response in MPT group was shorter than that in MP group (2 months vs 3.5 months，P<0.05).The incidence of grade Ⅰ-Ⅱ side effects in MPT group was
higher than that in MP group (50% vs 24%, P<0.05).However, there was no difference in the incidence of grade Ⅲ-Ⅳ side effects between two groups.Conclusion Compared with  MP regimen, the ORR of MPT is significantly increased, the time of response to treatment is markedly shorten.The incidence of adverse reactions is higher in MPT group, but there is no difference in the incidence of grade Ⅲ-Ⅳ side effects between MPT group and MP group, and the patients can tolerate  well.

Objective  To explore the efficacy and security of ballon kyphoplasty（BKP）in treatment for osteoporosis vertebral compression fractures, and to search for effective mthods to improve curative result and decrease complication.
Methods 51 patients were divided into BKP group and NSM group by patient’s will or general conditon, 26 patients were treated with ballon kyphoplasty(BKP group), 25 patients were treated with bed rest, analgesics and bracing（NSM group）.There were no differences in age, gender, VAS, BMD and severity of fracture between
two groups.15 and 18 patients were followed up for 6 months respectively in BKP and NSM groups, the VAS,vertebrae wedge angle and complications were observed and compared.Results In BKP group, the VAS were 1.6 and 2.7 in 6 h after operation and 6 months after operation,respectively.The Cobb angle were 11.6°and 16.3° in 6 h after operation and 6 months after operation,respectively.In NSM group, the VAS were 5.7 and the Cobb angle were 22.4° in 6 months after operation.Compared with NSM group, BKP group get better VAS, kyphosis reduction(P<0.05), without serious complications.Conclusion The treatment of BKP for osteoporosis vertebral compression fractures is effective and reliable.
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Objective  To compare  the clinical effects of extensive devascularization around the cardia plus cerclage on gastric fundus and extensive devascularization around the cardia  plus    application of staplers  on  esophagus transection for treatment of the  upper gastrointestinal hemorrhage of portal hypertension.Methods The clinical data  of 151 cases of upper gastrointestinal hemorrhage of portal hypertension with treatment of  portal azygous di
sconnection  were retrospectively analyzed,including 122 patients with extensive devascularization around
the cardia plus cerclage on gastric fundus  (gastric fundus cerclage group) and 29 patients with extensive devascularization around the cardia  plus    application of staplers  on  esophagus transection  (esophageal transection group).The  preoperative and    postoperative  liver function,ascites, hepaticencephalopathy,  astrointestinal bleeding,abdominal hemorrhage,gastrointestinal fistula rate of perioperative complications and long-term recurrent upper gastrointestinal bleeding rate of the patients in two groups were compared.
Results Compared with gastric fundus cerclage group,the more significant impact on patients with liver function and nutritional status,long hospital stay,the high incidence of postoperative gastrointestinal fistula were found in esophageal transection group.In comparative analysis of long-term postoperative bleeding, long-term rebleeding rate of the patients in esophageal transection group(variceal bleeding and  gastric mucosal lesions rebleeding) was 30.0% (15.0%,15.0%) which was higher than that in gastric fundus cerclage group (8.5%) (4.9%,3.6%).
Conclusion Compared with extensive devascularization around the cardia  plus    application of staplers  on  esophagus transection,extensive devascularization around the cardia plus cerclage on gastric fundus has the advantages on liver function,smaller and fewer complications,shorter hospital stay,and can effectively control
the long-term incidence of rebleeding.The operation is simple and easy to grasp.

Objective  To investigate the correlation of soft tissue structure of upper airway with the pathogenesis of obstructive sleep apnea syndrome (OSAS) in adolescents age group by analyzing magnetic resonance imaging (MRI) of upper airway.Methods  The subjects were divided into obese OSAS, obese controls and normal weight controls groups according to the results from polysomnography and body mass index measurements; Upper airway was scanned by MRI sagittally and axially; upper airway at all levels and soft tissue was analyzed by Amira Medical image analysis system.Results Tongue volumes in obese OSAS and obese controls were significantly greater than that in normal weight controls (P<0.05); tonsil and adenoid volumes in obese OSAS were significantly higher than those in two control groups (P<0.05 or P<0.001), but no significant difference was found between two control groups.The volumes of lateral pharyngeal wall in obese OSAS were higher than those in obese controls and normal weight controls (P<0.05 or P<0.001), and they were higher in obese controls compared with normal weight controls (P<0.05).In obese OSAS group, positive correlations were found between volumes of lateral pharyngeal wall and apnea/hypopnea index (AHI) (r = 0.879,P<0.01), as well as volumes of tonsils and AHI (r = 0.824, P<0.01).Conclusion  Obesity can increase the soft tissue volumes around upper airway, thereby increase the upper airway obstruction; lateral pharyngeal wall and adenoid volumes play major roles in evaluating the severity of OSAS in adolescents.
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Abstract:Objective To investigate and analyze the knowledge level and change of iodized salt coverage among population with iodine deficiency disorders (IDD) in Shandong Province before and after the interventions, and to provide the basis for the development of health education control strategies for IDD in Shandong Province. Methods Forty-five counties were randomly selected to conduct health education interventions among 100 counties with IDD in Shandong province.Three townships were randomly selected in sample counties, 30 villages (or communities) were randomly selected among sample townships, 30 students graded 4-6 and 15 housewives were randomly selected in each sample village or community.Questionnaires on IDD knowledge were conducted and household salt was collected to measure knowledge level and coverage rate among the subjects, respectively.Results The knowledge rate among school students was increased from 81.88% to 97.06%, 15.18% higher than that in baseline survey. The knowledge rate among housewives was increased from 82.59 to 95.46%, 12.97% higher than that in baseline survey.The coverage rate was increased from 98.95% to 00.00%.Conclusion IDD health education is an effective measurement.

Abstract:Objective To evaluate the prevalence,awareness rate,treatment rate,and control rate of hypertension and risk factors in Dehui city of Jilin Province,and provide reference to prevent hypertension for the middle-economy county-level city.
Methods A cross-sectional study was performed among 3 778 subjects in Dehui city of Jilin province.The subject underwent a standard questionnaire,biochemical tests and physical examinations.Logistic regression analysis was used to identify the risk factors of hypertension.ResultsThe prevalence of hypertension was 41.0% in this area;the awareness rate,treatment rate  and  control rate  of hypertension were 8.95%,6.38% and 0.45%,respectively. The awareness rates,treatment  rates and control rates of hypertension in city and rural areas were 9.9%,7.4%,0.4%； 7.6%,5.0%,0.6%;respectively.The prevalence of hypertension was 44.5% in city，and 36.3% in cural，there was significant differences between city and rural areas (P<0.05).Binary Logistic regression analysis indicated that the relative risks (95% confidence interval) of old age,male,centralobesity,drinking,famil
y history of hypertension,diabetes mellitus,dyslipidemia and work  for hypertension were 1.06 (1.05-1.06),1.56 (1.29-1.90), 2.42 (2.07-2.82),1.24 (1.01-1.52),1.95 (1.55-2.45) and 1.56 (1.17-2.10),1.62 (1.39-1.88),1.04 (1.01-1.08),respectively.Conclusion The prevalence of hypertension is higher in Dehui city of Jilin Province than other areas,but the  awareness rate,treatment rate and control rate are lower.
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Abstract:Objective  To explore the effects of caloric restriction and exercise on lipoprotein particle diameter and distribution of female obesity and provide evidence for obese patients to formulate special therapeutic schedule.
Methods Eighty female obese aged 30-60 years were randomly divided into control group (CG,n=20),diet group (DG,n=20),exercise group (EG,n=20) and diet+exercise group (DEG,n=20).The subjects in DG and EG performed diet and exercise interven
tion respectively and those in DEG simultaneously conducted two intervention style above,while those in CG maintained their routine lifestyle.The test duration was 8 weeks.The lipoprotein particle diameters were measured by polyacrylamide gel
electrophoresis before and after test. Fixation,staining and photograph were made after plasma electrophoresis.Standard curve was determined by taking natural logarithm of standard protein diameter as ordinate and migration lenth as abscissa.
The diameters of low density lipoprotein (LDL) and high density lipoprotein (HDL) particle  were figured out according migration lenth on gel board of the samples.Results Compared with before test,8 weeks after test,the LDL particle diameter was increased (P<0.05),the proportion of small low density lipoprotein (sLDL) particle was decreased (P<0.05) and the proportion of large low density lipoprotein (lLDL) particle was raised (P<0.05),the diameter and distribution of HDL had no significant change (P>0.05) in DG; the HDL particle diameter was elevated (P<0.01),the proportion of small high density lipoprotein (sHDL) particle was reduced (P<0.05) and the proportion of large high density lipoprotein (lHDL) particle was raised (P<0.05),the diameter and distribution of LDL had no significant change (P>0.05) in EG; both of the LDL and HDL particle diameters were  elevated (P<0.01 or P<0.05),the proportions of sLDL and sHDL particles were reduced (P<0.01) and the proportions of lLDL and lHDL particles were  raised (P<0.01) in DEG; all indices in CG showed no significant differences (P>0.05).Conclusion Caloric restriction or exercise could partly have impact on LDL and HDL particle diameters and distribution of female obesity while combination of caloric restriction and exercise may simultaneously improve lipoprotein particle diameter and distribution,thus reduce cardiovascular risk level of female obesity.
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