To investigate the expression of P53 protein in hippocampal gyrus in rats with Bcl-2 overexpression after global cerebral ischemia/reperfusion and discuss the effect and mechanism of anti-apoptosis of  Bcl-2.             Methods 90 healthy male SD rats were randomly divided into 3                           groups(n=30).Ischemia-reperfusion group(IR group): global cerebral ischemia-reperfusion model was produced by 4-VO method.Reperfusion were performed 6 min after  cerebral ischemia；sham operation group(SO group): the blood vessels were just exposed without occlusion；Bcl-2 overexpression group(Bcl-2 group):the rats were  dealed as the  same as IR  group after establishment of  the model of rats with Bcl-2 overexpression.All the animals were executed at 6,12,24,48,72 and 96 h by 4% poly-formaldehyde perfusion.Immunohistochemical staining,TUNEL staining and HE staining of brain tissue section were performed.The changes of neuron form of hippocampal gyrus,the expression of P53  in CA1 and CA3,and the number of apoptotic cells were observed with light microscope.All the data was analyzed by SPSS 10.0.Results ①24 h after global cerebral ischemia-reperfusion,the P53 protein expressed weakly in CA1,increased gradually and peaked at 72 h,then decreased;and mainly localized in cell nucleus.The  expression of P53 protein in CA3 appeared at 48 h after global cerebral ischemia-reperfusion,peaked at 72 h,which were weaker than that in CA1.The  expressions of P53 protein in CA1 and CA3  in Bcl-2 group were much weaker than those in IR group（P<0.05）.② HE staining showed the number of neuron in CA1 in IR group was decreased at 72 h after global cerebral ischemia-reperfusion,companying with neuron disarrangement,nucleus membrane indistinction and nucleolus disappearance.The change in CA3 was slighter than that in CA1 （P<0.05） and the changes  in Bcl-2 group was slighter than that in IR group（P<0.05）.③ TUNEL staining showed the number of apoptotic cells peaked at 24 h after global cerebral ischemia-reperfusion in CA1 of hippocampus（P<0.01）,continued until 72 h in IR group compared with SO group.While the positive cells appeared mainly at 72 h in CA3 compared with IR group.The number of apoptotic cells in  Bcl-2 group was less（P<0.05）.Conclusion Overexpression of Bcl-2 can inhibit  the expression of P53 in hippocampal gyrus  after global cerebral ischemia-reperfusion in rats.

To investigate the expressions of Ets family transcription factor ER81 in testis and epididymis of busulfan-treated mice,and explore its  function on the proliferation and differentiation of spermatogonia.Methods Busulfan peritoneal injection (10 mg•kg-1) was performed and mouse testis and epididymis were collected on the 0th,3rd,5th,8th,10th,18th days after injection, respectively.The mRNA expression levels of ER81 in samples were analyzed by semi-quantitative RT-PCR.Results Compared with the 0th day,the expression of ER81 in testis was significantly decreased on  the  5th day (P<0.01) and then recovered gradually.In epididymis,compared with the 0th day,the expression of ER81 on the 8th day was significantly decreased(P<0.05).Conclusion ER81 may modulate the differentiation of spermatogonia.

To check the effect of interferon regulatory factor 7 (IRF7) in the defense against the viral infection in hepatocytes by evaluating the function of IRF7 in the immune response of hepatocytes.Methods The human primary hepatocytes,immortalized human hepatocytes HuS-E/2 and hepatoma cell line HuH-7 cells were used for this study.After Sendai virus (SV) infection,the expressions  of IFNα1,IFNβ,IFNλ3,and RIG-I in hepatocytes and HuH-7 cells were detected by RT-PCR.The function of IRF7 in hepatocytes cells was blocked by transfection of the expressing plasmid of dominant negative form of IRF7 pcDNA3-7DN.The expressions of IFNα1,IFNβ,and IFNλ3  were detected by RT-PCR and real time PCR. The HuH-7 cells was infected with genotype 2a HCV JFH1 after transfection of the expressing plasmid  of IRF7 pcDNA3-IRF7.The infection efficiency was detected with indirect immunofluorescence.Results The induction of mRNA of IFNα1,IFNβ,IFNλ3,and RIG-I was not detected in HuH-7 cells after SV infection in 12 h,while that was detected in hepatocytes at the early stage (3 h).The expressions of the genes mentioned above were downregulated at the early stage of SV infection after the blockage of the function of IRF7,and the mRNA levels of these genes were significantly decreased in the first 3 h after SV infection in the IRF7 knockdown HuS-E/2 cells (p<0.001).The signal of infection and replication of JFH1 were　not detected in the HuH-7 cells with ectopic expression of IRF7.Conclusion The constitutively expressed IRF7 in hepatocytes is closely related to the immune response after infection,and plays an important role in the defense of viral infection.

Abstract:Objective
To study the effect of heterogeneous gene promoter on regulation of exogenous gene expression   in Jurkat cells and provide the theoretical basis for establishment of leukemia animal model.Methods The pig CD4 gene promoter was focasted and cloned by bioinformatics means;then using the cloned CD4 genen promoter,the lentivirus package was performed and the lentiviral vector was constructed.Affter the packaged virus was obtained the titer of virus was detected and the 293 T cells and Jurkat cells were infected to perform green fluorescent protein detection.The expressions  of exogenous genes of the pigs CD4 gene promoter and EF-1α promoter were compared.Results The pig CD4 gene promoter was successfully forecasted and the lentiviral vector was successfully constructed and the virus was packaged out.The dection of titer of virus and infection of 293 T cells and Jurkat cells showed that the start effect of GFP of two promoters in the cells were similar.Conclusion The pig CD4 gene promoter has not specific start of exogenous genes in leukemia Jurkat cells.The pig CD4 gene promoter can serve as a kind of different choice start of heterologous gene in leukemia Jurkat cells.

To construct the skeletal muscle-specific FSⅠ-Ⅰ eukaryotic expression vector and identify the integration of FSⅠ-Ⅰ gene in porcine embryonic fibroblasts,and to provide a basis for study on  the effect  of FSⅠ-Ⅰ on skeletal muscle.Methods The FS A (containing FS N and FSⅠ) and FSⅠ were amplified by RT-PCR from porcine skeletal muscle;the MCK enhancer,ACTA1 promoter and BGHpolyA were amplified by PCR from mouse genenome,human genenome and pcDNA3.1(+), respectively.The  four fragments mentioned above were subcloned into the eukaryotic expression vector pGKneotpAlox2,and then the vector was transfeced into porcine embryonic fibroblasts  using the FuGENER　HD reagent,according to the manufacturer’s protocol.The cells were selected with G418 antibiotic and identified by PCR amplification.Results The FS Ⅰ-Ⅰ skeletal muscle -specific expression vector was successfully constructed  and integrated to the genome of the porcine fibroblasts and  the clone cells integrating the FSⅠ-Ⅰ gene were obtained.Conclusion The porcine fibroblast clone  expressing FSⅠ-Ⅰ  is obtained and it should be of great value to the construction of FSⅠ-Ⅰ transgenic swine.

To study the effects of deoxycytidine kinase (DCK) on drug sensitivity and radiosensitivity of human cervical carcinoma cell line HeLa and to provide theoretical basis for gene therapy of cervical carcinoma. Methods FuGENE 6 liposome was used to transfect HeLa and establish RNAi models,the cells were divided into control,empty-vector and DCK-siRNA group,the expressions of DCK mRNA and protein before and after transfection were detected by quantitative real-time PCR and Western blotting,the abilities of proliferation were measured by cell counting,the drug sensitivity to cytarabine（AraC） was determined by MTT assay,the clonogenic formation test was used to assess the radiosensitivity.Results Compared with control group and empty-vector group, the    expressions of DCK mRNA and protein in siRNA group were significantly decreased (P<0.05).There was no significant difference of growth curve among three groups.The drug sensitivity to AraC in siRNA group was decreased,while the radiosensitivity was increased (P<0.05).Conclusion Inhibition of DCK could reduce the drug sensitivity and increase the radiosensitivity in human cervical carcinoma cells.

To construct the expression plasmid of pcDNA3.1myc-HisA-Smad2/3/4 and identify its fusion  protein expression.Methods pcDNA3.1- Smad2/3 and pGEX2T-Smad4 were used as templates,and the special primers were designed.The Smad2/3/4 coding sequence was amplified by polymerase chain reaction (PCR) method and subcloned into pcDNA3.1myc-HisA vector. After the target region was sequenced,the plasmid was transfected into HEK293 cell line. The expression of the recombinant plasmid in HEK293 cells was detected by Western blotting.Results Smad2/3/4 was constructed into expression vector pcDNA3.1myc-HisA successfully.The lengthes of the fragments were 1 401,1 275 and 1 656 bp,and they were identified by restriction enzymes digestion.The expression of pcDNA3.1myc-HisA-Smad2/3/4 fusion protein was proved by Western blotting.Conclusion The eukaryotic expression plasmid  pcDNA3.1myc-HisA-Smad2/3/4 is successfully constructed,and the expression of pcDNA3.1myc-HisA-Smad2/3/4 fusion protein is identified.

Abstract:Objective
To observe the changes of the orexin-B cells and their nerve fibers in the rat epilepsy model at different time points and elucidate their roles in the seizure.Methods Seizures were induced by intraperitoneal injection of kainic acid(KA),then the changes of the orexin-B immunoreactive cells and nerve fibers were detected by immunohistochemistry at 8 h,1 d, 3 d,7 d and chronic relapsing time after the termination of epilepsy respectively.Results The orexin-B immunoreactive cells mainly distributed in the hypothalamus and perifornical nuclei,not cerebral cortex and other brain tissues,but the suspicious positive cells were  detected in the hippocampus.The orexin-B immunoreactive nerve fibers  distributed widely.The orexin-B immunoreactive cells were decreased slightly in the first and  resumed，then were decreased and resumed again.And the cell count of orexin-B was significantly  decreased in 7 d group compared with other groups(P<0.05).The change trend of cell count of orexin-B was opposite to the one of nerve fibers. Conclusion The  orexin-B immunoreactive cells and the immunoreactive fibers of the orexin-B have changes in the process of  the seizures,it  shows that they may involved in the regulation of epilepsy.And it can help to control seizures by increasing the  concentration of orexin-B.

To explore the cell labeling ability of CM-Dil on bone marrwow mesenchymal stem cells (BMSCs) in vitro as well as its tracking capability in intestine tissues of burned rats.Methods BMSCs were isolated,purified and expanded by the method of adherent culture,and were induced to differentiate into neuron-like cells by basic fibroblast growth factor (bFGF),epidermal growth factor (EGF) and retinoic acid (RA).The surface markers of neuron-like cells differentiated from BMSCs,microtubule-associated protein-2 (MAP-2) and neuron specific nuclear protein (NeuN） were detected by immunocytochemistry.The BMSCs were labeled with CM-Dil at the doses of 4,6,8 mg•L^-1.Then the cytotoxic effect was determined by CCK-8 assay,the labeling rate was examined using flow cytometry.107 BMSCs labeled by CM-Dil were administrated by retro-orbital intravenous injection after burned models of Wistar rats were prepared.The intestine tissues were obtained and fixed on the 2 weeks and 6 months after the burning,then the engraftment of the labeled BMSCs were observed using a laser scanning confocal microscope.Results The nueon-like cells differentiated from BMSCs expressed neuron markers,NeuN and MAP-2. The results of CCK-8 assay showed that compared with control group,the cell viability in 8 mg•L^-1 group was significantly decreased (P<0.05）.There were no significantly differences between 4,6  μg•mL^-1groups and control group.No significant adverse effects on BMSCs were observed during CM-Dil labeling at the dose of 4 mg•L^-1,and the labeling rate was as high as 93.9%.The morphological changes of the BMSCs were not shown.Frozen sections of intestine tissues were performed on the 2 weeks and 6 months after burning,the CM-Dil labeled BMSCs were engrafted at the rat intestine tissues.Conclusion CM-Dil is capable of labeling BMSCs in vitro,thus could be utilized as tracking of BMSCs in the intestine tissue of the burned rats.

To explore the influences of low level hydroquinone (HQ) in biological characteristics and the expression of poly(ADP-ribose) polymerase-1(PARP-1)gene in rat bone mesenchymal stem cells(BMSCs),and elucidate the toxic effect of low dose HQ on bone marrow.Methods HQ dissolved in PBS buffer at the concentrations of 2.5,5.0 and 10.0 μmol•L^-1 were respectively given to BMSCs,and the cells treated with PBS only were used as control group.The cell viability and proliferation were detected with CCK-8 assay kit,the apoptosis of BMSCs  was measured  by Annexin V/PI apoptosis assay kit,while the PARP-1 gene expression was determined by reverse transcription real-time RT-PCR.Results Compared with PBS control group,the ability of cell proliferation raised obviously (P<0.05);the apoptotic rates of BMSCs in 5.0 and 10.0 μmol•L^-1  HQ groups 48 h after exposure were significantly decreased (P<0.05).The expressions of PARP-1  in 2.5,5.0,and 10.0 μmol•L^-1　HQ groups 48　h after exposure were 0.92,0.56(P<0.05),and 0.45 folds of control group(P<0.05).Conclusion Low level HQ could inhibit the apoptosis of BMSCs and PARP-1 gene expression,while promote the cell growth.

Abstract:Objective
To investigate the effects of activin A (ACTA) and  follistatin (FS) in alcoholic liver injury in mice,and  to clarify the mechanism of alcoholic liver disease.Methods Mouse models of alcoholic liver injury were established.24,72,120,168 h after establishment were used as the time points,the serum ACTA and FS levels and liver function changes  in 40 cases of alcoholic liver injury mice and 40 healthy mice were detected with ELISA method. The  liver tissues of model mice and healthy control mice were stained with  HE  and immunohistochemical method to observe the  histopathological changes of liver tissues and expressions of ACTA and  FS.Results The levels of  serum alanine aminotransferase (ALT),aspartate aminotransferase (AST) and ACTA in control group at each time point did not change significantly;while in model group,the  ALT,AST and ACTA levels  changed at different degrees,of which ALT,AST levels at  24 h for the highest,ACTA had  the highest level at  72 h.The  ALT,AST,and ACTA levels at the four time points in model group had significant differences   compared with the corresponding control group(P<0.01).At four time points, the FS levels  had no significant difference between model group and control group(P> 0.05).The control mouse liver tissue almost did not express  ACTA,but positively expressed  FS.ACTA highly expressed in the liver tissue around the portal area of model mice.There was no significant difference of  FS expression in liver tissues between control mice and model mice.Conclusion ACTA and FS  have  different changes in alcoholic liver injury,and ACTA and FS system imbalances may be the important reasons of alcoholic liver disease and  liver fibrosis.

To construct the genetic engineering bacteria highly expressing fusion protein of glutathione-S-transferase ( GST ) and C-terminal 71 amino acids of fibroblast growth factor 23 ( FGF23CTR ),and to test the immunogenicity of the fusion protein and  to provide experimental data for FGF23 specific monoclonal antibodies and the development of diagnosis reagent for  chronic kidney disease.Methods After FGF23CTR gene fragment was obtained by PCR,it was fused with GST by expression vector pGEX-4T-1,then was transformed in E.coli BL21(DE3). By inducing with 1IPTG,the fusion proteins were expressed solubly.The fused protein was purified by Glutathione Sepharose4B.The purity of GST-FGF23CTR by SDS-PAGE was shown to be higher than 90%.The   Balb/C mice  were immuned   with fusion protein to pepare antiserum,then  ELISA was used to assay antiserum titer.Results The GST-FGF23CTR expression vector was constructed successfully.The purity of fusion protein was more than 90% after purification,and it positively reacted  to the commercialization of FGF23 polyclonal antibody.The ELISA result showed that the confirmed fusion protein had  good immunogenicity.Conclusion The genetic engineering bacteria of GST-FGF23CTR is successfully constructed and the purified protein has good immunogenicity.The fused protein can be used for the FGF23 monoclonal antibody preparation and research on the biological function of FGF23.

Objective
To study the interaction between D29 and macrophages and observe the titer variation of D29 both inside and outside the Mycobacterium tuberculosis(MTB) after phagocytised by macrophages and provide references for study on the mechanism and clinic therapy dose for treating tuberculosis. Methods The macrophages cultured in vitro were cracked at different time after D29 was mixed to estimate the titers of D29,at the same time,the supernatant fluid was collected after coincubated for 12 h to detect the levels of NO and IL-12 by enzyme-linked immunosorbent assay(ELISA);the titers of D29 were calculated both inside and outside the MTB by fluorescence quantitative PCR at different time after coincubated with macrophages which had phagocytised MTB.
Results There were about 2.5×104PFU of D29 phagocytised by macrophages after coincubated with 1×108PFU for 20 min,many of them were inactive after 180 min,there were hardly active D29 at 190 min;the supernatant IL-12 and NO levels  had no significant differences between D29 group and blank control group(P>0.05),but they  were significantly lower than those in LPS positive group (P<0.05);D29 could enter MTB phagocytised by macrophages after coincubated for 30 min;when coincubated for 40 and 50 min,the titers of D29 were increased both inside and outside the MTB,then were decreased;they began to decrease largely at 190 and 210 min,while at 48 h D29 couldn’t be detected.Conclusion D29 might be inactive after phagocytised by macrophages,and D29 could not influence the immunologic function of macrophages;D29 can enter MTB phagocytised by macrophages,at first,with the prolongation of the time,the number of D29 in MTB is increased,afterwards,D29 begins to decrease and disappears in the end.

Abstract:Objective To investigate the effect of lancemaside D on the time effects of cell proliferation and cell cycle of HepG-2 cells and explore the anticancer active ingredients and mechanisms of codonopsis lanceolata. Methods The HepG-2 cells were treated with different concentrations (5,10,15,20 and 25 mg/L) of lancemaside D in vitro,meanwhile control group was set up. The proliferation inhibition of HepG-2 cells was detected by MTT assay,and the change of cell cycle  of HepG-2 cells was determined by flow cytometry. Results The results showed that the  inhibitory rate of cell proliferation of HepG-2 cells were  significantly higher than that in control group(P<0.01) after treated with 25 mg/Llancemaside D for 12,24,48 and 72 h;the cell numbers in lancemaside D groups were significantly smaller than that in control group after treated with lancemaside D for 48,72,96 and 120 h (P<0.05),among 5,15,20 and 25 mg/Lgroups there were significant differences (all P<0.05);the HepG-2 cell proliferation speed was delated with the increasing of lancemaside D concentration. The cell cycle detection results showed that in the lancemaside D concentration range of 10-25 mg/L,the percentages of HepG-2 cells at  G0/G1 phase were 55.97%±0.42%,57.16%±0.13%,57.48%±0.15% and 60.39%±0.32%, respectively,they were significantly higher than that in control group(53.22%±0.28%)(P<0.05). In the same concentration range of lancemaside D,the percentages of HepG-2 cells at S phage were 27.47%±1.42%,26.15%±2.71%,26.34%±0.67% and 24.81%±0.46%,they were significantly lower than that in control group(33.47%±0.25%)(P<0.05),which suggested a G0/G1 cell cycle arresting in lancemaside D group. Conclusion Lancemaside D strongly inhibits the growth of HepG-2 cells in vitro and furthermore cause cell DNA metabolism disturbance.

Objective To investigate the role of Ku70 in the chemoresistance of lung cancer and the potential of Ku70 small-interfering RNA (siRNA) as a therapy for reversal of cisplatin resistance in lung adenocarcinoma.Methods The Ku70 mRNA and protein expression levels were dectected by RT-PCR and Western blotting methods in human lung adenocarcinoma cells A549 and their cisplatin-resistant variant A549/DDP.The A549/DDP cells were divided into 3 groups:blank control,negative control group in which the cells were tranfected with nonspecific si-RNA（si-Scramble） and experiment group  in which the cells were transfected with Ku70-specific siRNA(si-Ku70).The cells  in negative control group and experiment group were transfected 48 h prior to treatment with cisplatin.The cell survival was assessed by MTT assay.The apoptotic rate was determined by  flow cytometry.The Caspase-3 activity was detected by spectrophotometer.Results  The Ku70 mRNA and protein expressions in A549/DDP cells were significantly elevated compared with the parental A549 cells（P<0.01）.The si-Ku70 down-regulated the mRNA and protein expressions of Ku70 in A549/DDP cells.After treated with different concentrations of cisplatin for 24 h,the cell viabilities of si-Ku70 cells were decreased significantly compared with  control group（P<0.01）.After treatment with 6 mg/l isplatin for 24 h,the apoptotic rates and the activites of Caspase-3 in experiment group were increased significantly compared with control group（P<0.01）.Conclusion The overexpression of Ku70 in cisplatin-resistant human lung adenocarcinoma cells suggests that Ku70 might be involved in the chemoresistance of lung cancer.Ku70-specific siRNA can down-regulate the expression of Ku70,promte apoptosis and reverse resistance to cisplatin in cisplatin-resistant A549/DDP cells.Ku70 may be a new target for reversal of chemoresistance of lung cancer.

Objective To investigate the interventional effect of oxymatrine (OMT) on focal cerebral ischemia and explore the mechanism of its intervention on oxidative stress response.Methods The Wistar rats were randomly divided into sham operation group,model group,positive drug group,OMT1 (35 mg/kg) group,OMT2 (70 mg/kg)　 group and OMT3 (105 mg/kg) group (n=10).The drug was continuously administrated 5 d before the surgery of middle cerebral artery occlusion (MCAO) was carried out.The neurological score was recorded 24 h after operation and the  morphological changes of ischemic brain tissue were observed by HE staining.The interventional effects of OMT on oxidative stress of cerebral ischemic rats were investigated by measuring the activities of superoxide dismutase (SOD),catalase (CAT) and glutathione-SH peroxydase (GSH-Px) and the content of malondialdehyde (MDA) in peripheral serum.Results Compared with model group,the neurological scores of rats in OMT groups were significantly decreased(P<0.05 or P<0.01),the morphological characteristics of ischemic brain tissue were maintained and  the damage of tissue  was alleviated with a certain degree;the  activities  of SOD,CAT and GSH-Px were increased (P<0.05 or P<0.01)　and  the contents of MDA reduced (P<0.05 or P<0.01)  with a  dose-effect relationship.Conclusion OMT can intervene the focal cerebral ischemic injury by influencing the oxidative stress respone in rats with cerebral ischemic injury.

Objective To investigate the inhibitory effect of low-dose lidamycin on the growth of P19 cells,in order to provide a reliable evidence for targeting cancer stem cells in cancer therapy.Methods  MTT assay was used to measure the dose curve and time curve after P19 cells were treated with lidamycin.Flow cytometry was employed to examine the  apoptosis and cell cycle of P19 cells treated with lidamycin and adriamycin.The  Oct4 expressions at mRNA and protein levels were tested by Real-time PCR,Western blotting and immunocytochemistry when the P19 cells were exposed to lidamycin and adriamycin.Results After exposure of the cells to lidamycin and adiramycin,the IC^₅₀ of lidamycin and adiramycin in P19 cells were 1×10^－11 mol/L and 0.5×10^－9 mol/L,respectively.Low doses of lidamycin range from 0.001 to 0.1 nmol/L^-1 were still effective to inhibit P19 embryonal carcinoma cell growth in 120 h. Flow cytometry indicated that  low-dose  lidamycin induced G1 stage arrest in P19 cells without apoptosis and  adiamycin induced G2 stage arrest.Real-time-PCR,Western blotting and immunochemistry showed that the expression levels of Oct4 were decreased in all test when the P19 cells were exposed to lidamycin,but not to adiamycin.Conclusion The inhibitory effect of low-dose lidamycin on P19 cell growth may be related to down-regulation of embryonic stem cell-like gene Oct4.

Objective To study the effects of telomerase inhibitor zidovudine (AZT) on the cell morphology,survival rate and telomerase activity of osteosarcoma cell line (HOS) in vitro,and to provide the experimental basis for the application of telmoerase inhibitors in  treatment of osteosarcoma. Methods The osteosarcoma cells (HOS) at logarithmic growth phase were divided into  AZT groups and control,the  medium were added with  100 μL AZT diluted with culture medium  with different  concentrations (1，10,100,1000  μmol/L)  and the same volume of culture medium,24,48,72 h after culture,the morphological changes of osteosarcoma cells were observed with light microscope;the survival rate was detected by MTT assay;the changes of telomerase activity were determined by TRAP-PCR-ELISA.Results After treated with  AZT,osteosarcoma cell line (HOS) had an obvious change in shape by size decreasing and shrinkage.The short spindle became polygonal or irregular in shape,and with neighboring cells detachment.Compared with control group,the survial rates in AZT groups were significantly decreased (P<0.05) in a time- and dose- dependent manner.Its IC50 was 100 μmol/L.The telomerase activity of HOS cells was decreased by 55.74％after treated with 100 μmol/L AZT for 72 h.Conclusion AZT has a significantly inhibitory effect on the proliferation of HOS cells,and AZT can reduce the telomerase activity of osteosarcoma cells.

Objective To observe the cell count changes of A549 cell line and human embryonic lung diploid cell line 2BS after irradiated by ¹²⁵I seeds with different doses,and to study the growth inhibition of ¹²⁵ I on  this two kinds of cell lines，and to determine its clinical safety dose in treatment of  non-small cell lung.Methods ¹²⁵I seeds with different doses （low dose:0.2 mCi,mediate dose:0.4 mCi, high dose:0.8 mCi）were chosen and  put  into A549 cells  and human embryonic lung diploid cell line 2BS  in vitro,the  cells on the 2nd,4th,6th and 8th days after irradiation  were collected, the alive  cells were counted by cells dyeing experiments,then  the growth curves were drawn,and  the IC50 of the radioactive ¹²⁵I seeds to both two  cell lines were calculated.Results Compared with  blank and control groups，the cell proliferation trend of A549 cells in low dose group was not significantly influenced（P＞0.05）,but the growth  of A549 cells in mediate and high dose groups were inhibited in a time-dependent manner,there were   significant differences(P<0.05),
the most obvious change was on  the 6th day.The IC₅₀ of the radioactive 125I seeds to A549 cells was about 0.4 mCi.While the growth inhibition of ¹²⁵I 2BS  had no statistically significant differences between various dose groups(P>0. 05),and the IC₅₀ of the radioactive ¹²⁵I seeds to 2BS cell line was about 1.65 mCi.Conclusion 0.4 mCi of radioactive ¹²⁵I seeds  has already had the obvious damage effect on A549 cells,0.8 mCi of radioactive ¹²⁵I seeds has the   stronger effect.The IC₅₀ of the radioactive ¹²⁵I seeds to 2BS cells is about 1.65 mCi,so the clinical safety dosage is 0.4-0.8 mCi.

Objective To study the influence of oxytocin(OT) in the proliferation and osteogenesis activity of MG-63 cells,and to clarify its function on bone metabolism.Methods The MG-63 cells were incubated with  10,50 and  100 nmol/LOT for 1,3,5 and 7 d. The cell proliferation was detected by MTT,then  the minimum effective concentration was used to  detect the change of alkaline phosphatase (ATP) activity .Results The  MTT test results showed that    the  proliferation  of MG-63 cells after treated with  50,100 nmol/L OT for 1,3,5 and 7 d was  significantly higher than those in control group(P<0.05);and  the proliferation of  MG-63  cells after treated with 10 nmol/LOT for 7 d was significantly higher than that in control group (P<0.05).The ELISA test results showed that the ALP  activities of MG-63 cells after treated with 50 nmol/L  OT  for 3,5 and 7 d were sifnificantly higher than those in control group(P<0.05).Conclusion OT has a regulation effect on proliferation of MG-63 cells,and it can promote the  proliferation and differiation of MG-63 cells.

Objective To construct the eukaryotic expression plasmid pIRES2-EGFP-hVEGF165 of  human vascular endothelial growth factor 165(hVEGF165),and to transfect pIRES2-EGFP-hVEGF165 into human placental mesenchymal stem cells (HPMSCs) through liposome,and to identify the express activity of hVEGF165 and the multi- differentiation potential of HPMSCs  carrying the target gene. Methods From human leukemia cells HL-60 the gene fragment of hVEGF165 was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR),the pIRES2-EGFP-hVEGF165 recombinant plasmid was constructed,and its correctness was detected by restriction enzyme digestion;the activity of transcription and expression of  HPMSCs  transfected by liposome were respectively detected with RT-PCR,Western blotting and MTT assay; the enhanced green fluorescence protein(EGFP) was observed under fluorescence microscope;and the conditions of transfected target cells of empty vector containing reporter gene EGFP,eukaryotic expression vector carrying reporter gene and target gene were detected;and the multi-differentiation potential of HPMSCs after transfection was indentified.Results The recombinant plasmid pIRES2-EGFP-hVEGF165  was successfully constructed through enzyme cutting identification.The constructed plasmid of HPMSCs after transfection with pIRES2-EGFP-hVEGF165 had transcription and expression activities;the EGFP expression was detected  under fluorescence microscope,and the target gene was successfully transferred into target cells,and the HPMSCs carrying the target gene still maintained multipotentiality.Conclusion The eukaryotic expression plasmid pIRES2-EGFP-hVEGF165 carrying hVEGF165 with expression activity is successfully constructed,and the plasmid is successfully transferred into HPMSCs.The hVEGF165 in HPMSCs has the activities of transcription and expression,and hVEGF165 could promote the proliferation of HPMSCs,and the HPMSCs itself might have the endocrine function of hVEGF165.

Objective To observe the effects of 4,5,6,7-tetrabromobenzotriazole(TBB) on the expressions of CK2β and Akt in thyroid squamous cell carcinoma cell line (SW579) and to explore their mechanisms.Methods SW579 cells were cultured in vitro and divided into con
trol group,DMSO group，TBB (25 μmol/L) group， and Wortmannin (100 nmol/l group randomly.The inhibitory rate of the growth of SW579 cells was detected with MTT and the protein kinase activity was measured.The CK2β and Akt mRNA and protein expression levels were determined by RT-PCR and Western blotting.Results Compared with control group,the inhibitory rate of growth of SW579 in TBB group was increased（P<0.01） and the cellular activity of CK2 was decreased（P<0.01）. Compared with control group,the expression levels of CK2β mRNA and protein  in TBB group were significantly reduced（P<0.01）,while Akt mRNA and protein levels were unchanged（P>0.05）,but phosphorylation status of Thr308 and Ser473 of Akt  in TBB group was inhibited（P<0.01）.Conclusion TBB can inhibit the expression and activity of CK2β and down-regulate the activity of Akt of SW579 cells.

Abstract:Objective To study the inhibitory effects of tetraiodothyroacetic acid(Tetrac) on proliferation  of doxorubicin(Dox)-resistant human gastric cancer cell line SGC-7901/R in vitro and in vivo　and their action mechanisms.Methods Drug-sensitive(SGC-7901) and -resistant(SGC-7901/R) cells were cultivated  with different  concentrations of Dox or Tetrac for  4 d.The cell viability  was tested by MTT assay.Western blotting was used to detect the  expressions of P-gp,GST and SOD in drug-sensitive and drug-resistant  SGC-7901 cells.Dox-resistant SGC-7901/R cells were treated  with Tetrac either alone or in combination with each of Dox,etoposide (Etop),cisplatin (Cisp) 4 d later;the  cell viability,aging and apoptosis were  determined by  MTT assay,senescence-associated beta galactosidase (SA-b-Gal) and Hoechst staining.Nude mice were injected with Dox-resistant SGC-7901/R cells (10⁶/100  μL) and divided into 4 groups (n=7):saline control group,Tetrac (30 mg/kg^-1) group,Dox (2 mg/kg)  group and Tetrac (30 mg/kg) +Dox (2 mg/kg)  group.The growth of tumor in various groups was measured.Results The necrosis of two cell lines was significantly increased  with  the increasing of Tetrac concentrations,and all of them were dead  at 100 mg/L.The expression of drug resistance molecule P-gp was detected in SGC-7901/R cell line.The survival rates of SGC-7901/R cells after treated with Tetrac combined with each of Dox,Etop,Cisp or not were significantly decreased(P<0.001);the aging and death of SGC-7901/R cells after treated with Dox and Tetrac were significantly increased compared with using drug alone (P<0.05).In vivo study showed that  the growth of SGC-7901/R cells could be inhibited with Dox and Tectrac.Conclusion Tetrac can promote the aging and apoptosis of SGC-790/R cells by reducing the expression of drug resistance molecule P-gp, Tetrac is an effective chemotherapeutic agent in Dox-resistant cells.

Objective
 To prepare nanofiber dressing by using electrospinning technology and determine its bacteriostatic efficacy. Methods Polyacrylonitrile (PAN) was used for precursor,by applying electrostatic spinning technology and high temperature carbonization method carbon nanofibers were prepared.Useing silver mirror reaction,nanoscale silver / carbon nanofiber composite dressing was prepared.According to Kirby-Baucer method,antibacterial extrasomatic test was conducted with these two samples and gentamicin scraps of paper and urgotul.Results The composite dressing had nanofiber scaffold structure and carried anoscale silver.The in  vitro antibacterial experiment results showed that the nanoscale silver / carbon nanofiber composite dressing and urgotul had good antimicrobial effects on  Staphylococcus aureus(Pae),E.coli(Eco) and Pseudomonas aeruginosa(Sau);the effects of composite dressing were  better than urgotul on Pae and Sau,but was poor on Eco(P<0.05);the carbon nanofiber dressing had no antibacterial property.Conclusion The nanoscale silver / carbon nanofiber composite dressing with  nanofiber scaffold structure and good antimicrobial effect is a good burn wound dressing.

Objective
To study the protective effect of Lubao Health cultivation Drink (LHD) on overtiredness rats induced by simulating swimming overtraining and investigate its mechanism.  Methods The fatigue model was induced by simulating swimming overtraining for 8 weeks in rats. Male Wistar rats were divided randomly into 6 groups,including control group,general intensive swimming training group,overload swimming model group and low dose LHD group,middle dose LHD group  and high dose LHD group. The rats in A,B and C groups were treated with  (with 10mL?kg-1?d-1 i.g).  The rats in D,E and F groups were treated with different concentrations of LHD distilled water (10 mL?kg-1?d-1 i.g). After 8 weeks,the changes of body weight,urine protein (UP),blood sugar(BG),hemoglobin(Hb),serum testosterone(ST) and plasma corticosterone(PC) contents in rats were determined. The contents of dopamine(DA),noradrenaline(NA),5-hydroxy tryptamine(5-HT) and γ-Aminobutyric acid(GABA) in hypothalamus were determined with sandwich ELISA method,and the ratios of DA/5-HT,NA/5-HT and DA/NA were calculated. Results Compared with overload swimming model group,in middle and high dose LHD groups the body weight loss induced by simulating swimming overtraining from 5 to 8 weeks wwas significantly improved and the UP contents were decrease (P<0.05 or P<0.01),but BG,Hb,ST and PC contents had no significant changes.In addition,in middle and high dose LHD groups the contents of 5-HT were decreased,and the content of DA and ratios of DA/5-HT,NA/5-HT and DA/NA in hypothalamus were increased (P<0.05 or P<0.01). Conclusion LHD has protective effects on overtiredness rats induced by swimming overtraining through decreasing the content of 5-HT,and increasing the contents of DA and GABA and ratios of DA/5-HT,NA/5-HT and DA/NA.

Objective
To explore the effect of rosiglitazone in the treatment of Alzheimer’s disease(AD) and provide guidance for AD treatment. Methods A thorough literature search related to randomized controlled trial(RCT) or clinical controlled trial(CCT) on treatment of AD with rosiglitazone was performed among Medline and database of CNKI in accordance with the requirements of Cochrane systematic review. Two reviewers reviewed the original articles,abstracted data and assigned an initial quality rating independently. With RevMan 5.0 software,Meta-analysis was carried out on the three literatures which were accorded with the criterion. Results Eight studies were included. All the studies reported and measured the changes in ADAS-cog score from baseline to endpoint for active treatment compared with placebo and there was a significant difference between two groups（WMD=－0.39，95%CI=－0.69－－0.08,P=0.01）. No significant difference of safety was detected between placebo and treatment groups（OR=1.16，95%CI＝0.97－1.37，P=0.10）. But a significant difference was observed when the data was stratified by dosage,the incidence of adverse events in patients who took 8 mg rosiglitazone everyday was roughly as 1.33 times as those who took placebo（OR95%CI＝1.13－1.56）. Conclusion Rosiglitazone can to some extent improve cognitive function of AD patients. However,the safety should be considered when it is applied.
 

Objective
To detect the hMOF protein expression in renal cell carcinoma(RCC) tissue,and explore its relationship with the cliniopathological characteristics of kidney carcinoma.Methods Immunohistochemistry and qRT-PCR were used  to detect the hMOF expression in the normal kidney tissues and renal cell carcinoma tissues.The relationship between the hMOF expression and clinical features of kidney carcinoma  was analyzed.Results The　 qRT-PCR results showed that there were 4 cases in 9 cases of RCC (44.4%) which hMOF mRNA levels were decreased significantly.The immunohistochemistry results showed that there were 39 cases (62.1%,39/66) of RCC  which hMOF protein expression was negative or weakly positive,only 4 cases (6.1%,4/66) of normal kidney tissues which hMOF protein expression was negative or weakly positive,there was significant difference between two groups (P<0.05). The hMOF expression  in Fuhrman Ⅲ kidney carcinoma tissues  was significantly lower than that in Fuhrman Ⅰ-Ⅱ(P<0.05),and it was significantly lower in poorly and moderately differentiated RCC tissues  than that in well differentiated group(P<0.05).In addition,in hMOF lower expression group,the risk of lymphonode and distant metastasis was increased.Conclusion With the increasing of Fuhrman grade,the expression of hMOF protein in kidney carcinoma  tissue is decreased significantly.The hMOF protein may be associated with the occurrence and development of kidney carcinoma,and malignant degree.

Objective
To study the expressions of epidermal growth factor (EGFR) and transforming growth factor-α(TGF-α) in human prostatic adenocarcinoma (PAC),high grade prostate intraepithelial neoplasia (HGPIN) and benign prostatic hyperplasia (BPH) tissues,and to explain the effect of the expressions in the  development of  PAC.Methods 41 specimens of PAC  and specimens of BPH and 32 specimens  of  HGPIN were selected,the expressions of EGFR and TGF-α  were determined by immuohistochemisitry staining.The relationship between the expressions of EGFR and TGF-α  and  histopathologic classification was analyzed.Results ①The EGFR positive rates were 56.6%,53.7% and 85.4% in PAC,HGPIN and BPH，the differences between BPH and HGPIN,BPH and PAC were statistically significant（P<0.01）. There was no significant difference in  EGFR positive rate between PAC and  HGPIN. The EGFR positive rates were 53.9%,55.6% and 52.6% in well-differentiated,moderately differentiated and poorly differentiated PAC,the expressions of EGFR had no correlation with different Gleason grades in PAC （P>0.05）.②The TGF-α  positive rates in PAC and BPH were 56.1% and 85.4%,there was significant difference between them（P<0.01）.The TGF-α  positive rate were 53.8%,66.7% and 52.6% in well differentiated,moderately differentiated and poorly differentiated PAC,the expressions of TGF-α had no correlation with different Gleason grades in PAC （P>0.01）. The TGF-α positive rate in HGPIN was 56.3%,there was no significant difference compared with  PAC（P>0.05）,but there was significant difference compared with  BPH (P<0.01).The collective positive rate of EGFR and TGF-α  was 81.8%,the respective positive rate of  EGFR was 18.2% and the respective positive rate of TGF-α was 26.3% in PAC,they had  positive correlation（r=0.558，P<0.001） and they had no  correlation in HGPIN （r=0.111，P=5.450） and BPH （r=0.024,P=0.883）. Conclusion The EGFR and TGF-α expression levels in PAC,HGPIN and BPH are different,combined detection of them can be useful for the diagnosis,differential diagnosis and evaluation in benign and malignant prostate disease.The expressions of EGFR and TGF-α  play centain roles in the pathogenesis of prostatic adenocarcinoma．

Objective To observe the expression  characteristics of  cytoketatin19（CK19） and Galectine-3  in thyroid  puncture samples and analyze their applicantion in differential diagnosis for   benign and malignant thyroid disorders,and to explore the feasibility of application of Galectine-3 and CK19 in pathological diagnosis of small puncture samples.Methods The puncture tissues from 21 cases of papillary thyroid carcinoma(PTC) and 16 cases of benign thyroid lesions(BTL) were examined by HE section and the  expressions of Galectin-3,CK19 and Ki67 were detected by EnVision immunohistochemistry. The different expressions of the three proteins were compared between two groups and their roles in the pathological diagnosis in the puncture samples before  thyroid operation were analyzed.Results The positive rates of Galectin-3,CK19 and Ki67 in PTC group was were 90%（18/20）,95%（19/20）,and 60%（12/20），respectively.The positive rates of the three proteins in BTL group were 12.5%（2/16，mildly positve）,31.25%（5/16，mildly positive）, and 62.5%（10/16）,respectively. There were significant differences of Galectin-3 and CK19 between PTC and BTL groups（P<0.01）.There was no significant difference of Ki67 expression between PTC and BTL groups（P>0.05）.Conclusion The expressions of Galectin-3 and CK19 are significantly different between PTC and BTL.Combined detection of these two proteins plays an important role in differential diagnosis of PTC and can improve the diagnosis rate of puncture tissue of thyroid disorders before operation.
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Objective To observe the expressions of Snail and E-cadherin in laryngeal squamous cell carcinoma(LSCC) tissues and normal squamous epidermis tissues near cancer,and to clarify the relationship with the occurrence and development of LSCC.Methods Streptavidin peroxidase(S-P method) immunohistochemistry staining was used to detect the expressions of Snail and E-cadherin in 46 samples of LSCC and 46 samples of normal squamous epidermis near cancer.The relationship between the expressions of Snail and E-cadherin and the clinical pathological characteristics were analyzed.And the relevance of the expressions of Snail and E-cadherin in LSCC was also analyzed.Results ① The positive rates of Snail expressions in LSCC and  normal squamous epidermis near cancer were 69.57%(32/46) and 36.96%(17/46). The expression of Snail in LSCC was significantly higher than that in normal squamous epidermis near cancer (P<0.05).There was  significant correlation between the expression of Snail in LSCC and the differentiation and cervical lymph node matastasis (P<0.05).②The positive rates of E-cadherin expressions in LSCC and normal squamous epidermis near cancer were 47.83％(22/46)and 78.30％(36/46).The expression of E-cadherin in LSCC was significantly lower than that in normal squamous epidermis near cancer (P<0.05).There was significant correlation between the expression of E-cadherin in LSCC with the T stage,clinical stage,cervical lymph node matastasis and differentiation (P<0.05).③The expression of Snail in LSCC tissues was negatively correlated with the expression of E-cadherin (r=－0.569 0,P<0.01).Conclusion The expressions of Snail and E-cadherin  may be  associated with the occurrence and development of LSCC,and they could be used as the parameters which predict the  malignant degree  and  prognosis of LSCC.

Objective To study the expressions of heat shock protein 70(HSP 70) and its upstream regulation factor HSF1 in laryngeal papilloma and vocal leukoplakia  and clarify their action in the genesis of human laryngeal  carcinoma.Methods The expressions of HSP70 and HSF1 proteins were detected by  immunohistochemical method in 6 specimens with vocal leukoplakia (vocal leukoplakia group),6 specimens with laryngeal papilloma (laryngeal papilloma group ) and 7 specimens of vocal cord polyp (control group).Results The expressions of HSP70 and HSF1 in laryngeal papilloma and vocal leukoplakia  were higher than that in vocal cord polyp,the differences were significant(P<0.05 or P<0.01),the expressions of HSP and HSF1  in vocal leukoplakia  were the highest.Linear regression analysis of HSP70 protein and HSF1 expressions in vocal cord polyp,laryngeal papilloma and vocal leukoplakia  showed positive linear correlation (r=0.867,P<0.01 ).Conclusion The overexpression of HSP70 and HSF1 protein in vocal leukoplakia  may contribute to the genesis of human laryngeal  carcinoma.HSP70 and HSF1 may be used as indicators for early diagnosis of laryngeal carcinoma.

Objective To discuss the incidence of memory impairment in chronic obstructive pulmonary disease（COPD） in elderly patients and to provide the basis for the intervention of COPD.  Methods 138 elderly patients with COPD were selected as case group,90 cases of healthy elderly were used as control group.The second edition of Applied Behavioral Memory Test (RBMT-Ⅱ) on memory function was  used for personnel evaluation in two groups,including a total of 13 items,and the results were analyzed. Results Among 138 cases of COPD patients,there are  0 case of patients with no memory impairment(0%), 10 patients with mild memory impairment (7.2%),86 patients with moderate memory impairment (62.3%),42 patients with severe memory impairment (30.4%),the morbidity rate of memory impairment was 100%. In control group,there were 12 cases of patients with  no memory impairment   (13.3%), 43 patients with mild memory impairment (47.8%),29  patients with moderate memory impairment(32.2%), 6 patients with severe memory impairment(6.7%).There was statistically significant difference in morbidity rate between two groups (P<0.01).Individual preliminary scores,recalling the name,hidden goods,picture recognition,immediate story recall,delayed story recall,face recognition,immediate  letter recall,delayed letter recall,orientation,date and the total standard score  10 items in case group were significantly lower than those in control group,there were statistically significant differences (P<0.05 or P<0.01).Conclusion The elderly patients with COPD always have memory disorders，mostly moderate to severe,the intervention for stable period of   disease should be strengthened to  reduce the incidence of  memory disorders and  improve the    life quality of patients.

Objective
To study the effectiveness of dexmedetomidine combined with fibreoptic bronchoscope for tracheal intubation in difficult airways caused by huge goiter.Methods Thirty patients with  anticipated difficult airways caused by huge goiter were enrolled and randomly divided into dexmedetomidine group (n=15) and propofol group (n=15).The patients in dexmedetomidine group received a loading dose of dexmedetomidine (1.0 μg?kg-1),infused over 10 min,then pumped at continuous rate of 0.4 μg.kg^-1.h^-1.The patients in propofol group received a loading dose of 2.0 mg.kg^-1 and pumped at continuous rate of 5-8  mg.kg^-1.h^-1. The intubating conditions  were graded by a scoring system; the reactions to intubation such as coughing and patient tolerance were observed;the heart rates and mean arterial blood pressures (MABP) at different time points of baseline(T0),Rassay score 4(T1),intubating(T2),1 min after intubation (T3) and 3 min after intubation (T4) were recorded;adverse events and haemodynamic support were observed.Results  All the  patients in two groups were performed successfully with fibreoptic intubation.The patients in dexmedetomidine group  could keep better spontaneous breathing without respiratory depression,and were able to command and cooperate tracheal intubation,while in propofol group 11 patients (73.3%) could not cooperate tracheal intubation (P<0.05).With respect to intubation scores in propofol group,there were 7 cases of  vocal cord opening and 3 cases of  vocal cord movement;while in dexmedetomidine group,there were 12 cases in vocal cord opening and 3 cases in vocal cord moving.Compared with propofol group,the patients in dexmedetomidine group had more favorable intubation scores of  vocal cord movement (P<0.05).With respect to no reaction or slight grimacing of reaction to intubation comfort score,there were 8 cases in propofol group and 12 cases in dexmedetomidine group,and there was significant difference (P<0.05).The patients in dexmedetomidine group experienced fewer airway events and less heart rate response to intubation than those in  propofol group (P<0.05).Conclusion Compared with propofol in management of difficult airways caused by huge goiter,dexmedetomidine has better tolerance,and preserves a patient airway,and has more stable haemodynamic response to intubation.
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Objective To investigate both the short-term effect and side effects of anti-thymocyte  globulin (ATG) combined with cyclosporin A(CsA) therapy in patients with severe aplastic anemia(SAA) and provide basis for the treatment of SAA. Methods From December 2009 to May 2011,15 SAA patients treated by ATG combined with CsA therapy in our hospital were selected,the clinical effectiveness,the improvement of their symptoms after  treatment,the changes of peripheral blood cell counts and the incidence of different side effects in treatment period were observed.Results Among 15 patients,there were 10 patients reached the treatment assessment time point: 4 patients got　complete response(40.0%),4 got partial response(40.0%),while 1 patient had no　response(10.0%),1 patient died after ATG treatment(10.0%),the overall effective rate was 80.0%.The patients who responsed to this therapy got their blood cells recovery 1-2 months after treatment: the white blood cell counts and neutrophil counts recovered first,while the hemoglobin and platelet recovered at least 3 months after ATG treatment.During the ATG treatment,the most common side effect was varying degrees of acute allergic reaction and serum disease(53%),followed by infection(40%).Conclusion For the SAA patients who are not suitable for homatopoietic stem cell transplantation,ATG combined with CsA is a standard therapy,with a satisfactory curative effect.
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Objective To investigate the clinical efficacy of retroperitoneal laparoscopic surgery of adrenal pheochromocytoma by comparing the differences between open and retroperitoneal laparoscopic surgery of adrenal pheochromocytoma.Methods Based on patients’ requirements and surgeon’s experience,48 patients with  unilateral adrenal pheochromocytoma  were divided into  retroperitoneal laparoscopic group (n=23) and  open surgery group (n=25).The clinical efficacies in two groups were compared and  the differences between them in operation time,blood loss,intraoperative blood pressure control,postoperative hospitalization time and the incidence of postoperative complications were investigated.Results The operation in two  groups was successful.In retroperitoneal laparoscopic group and open surgery group,the  operation time were (60±25) min and (100±40) min,the blood loss were (42±16) mL and (156±45) mL,the blood pressure in operation  were(160±10) mmHg and (165±55) mmHg,the postoperative hospitalization time  were (5.2±1.2)d and (7.4±1.6) d.there were significant differences between two groups（all P<0.01）.There were no major postoperative complications.All the 48 cases were pathologically diagnosed as pheochromocytoma.All patients were followed up for 6-48 months,their clinical  symptoms disappeared.No tumor recurrences developed.Conclusion Retroperitoneal laparoscopic surgery is superior to open surgery in treating adrenal pheochromocytoma patients,which has less bleeding,less trauma and quicker recovery.It is therefore a safe and effective method in treating pheochromocytoma.

Abstract:Objective To study the clinical effects of high-frequency electric knife,CO₂ laser,microwave technique and conventional surgery in treating gingival hyperplasia caused by  ill porcelain-fused-to-metal (PFM) and explore the values of their chinical applications.Methods Different types and various degrees of 148 patients with gingival hyperplasia were randomly divided into  high-frequency electric knife group,microwave technique group,CO₂  laser group,and conventional surgery group,37 cases in each group.The patients were treated with the 4 kinds of methods mentioned  above.Then gingiva was removed and  its form was repaired to restore and reconstruct the periodontal physiological function and appearance.Results ①Compared with  conventional surgery group,in high-frequency electric knife,CO₂  laser,microwave technigue groups,the  operative time was shorher,the intraoperative blood loss was lower,the wound healing time was shortened,and the wound infection rates were decreased(P<0.05 or P<0.01).②The gingival indexes(GI),sulcus bleeding indexes(SBI) and modified bleeding indexes(mBI) in 4 groups after operation were significantly decreased compared with before operation(P<0.01).The GI in high-frequency electric knife group,CO₂  laser group,and microwave technique groups after operation were significntaly  lower than that in conventional surgery group (P<0.05).③ The one-year  efficiencies in high-frequency electric knife group,CO₂  laser group,and  microwave technique group were 97.3%,94.6%,and 97.3%,there were no significant differences between three groups (χ2 = 0.34,P> 0.05);but they were sifnificantly higher than that in conventional surgery group  (89.2%) (χ2 = 0.54,P<0.05).Conclusion High-frequency electric knife,CO₂ laser,and microwave technique   can be effective in the treatment of gingival hyperplasia,and superior to conventional surgery.

Abstract:Objective To analyze the feasibility of miniscrew placement in the buccal bone around maxillary molar roots for whole dentition distalization.Methods Pretreatment cone beam computed tomography (CBCT) images of 88 patients were evaluated,which were divided into two groups according to the number of the buccal root of maxillary second molars.The actual buccal bone thickness around the buccal roots of maxillary molars and bone thickness necessary for miniscrew placement there with the angle of 70° to the maxillary occlusal plane were measured.Upon comparison of the  two  sets data in each  group,the percentages of safety placement in patients were calculated.The interradicular space along the arch was measured for analysis of the available amount of distalization.Results In all the buccal areas there were certain percentages of patients could be inserted miniscrews safely.In the group of patients with two-buccal-root maxillary second molars,the highest safety rate of 85.70%,was around the mesiobuccal root of maxillary second molars;in the group of patients with single-buccal-root or fused root maxillary second molars,the highest safety rate of 91.38%,was around the maxillary second molar roots.The interradicular space along the arch in front of the mesial buccal roots was larger,and that in front of the distal buccal roots was smaller.Conclusion In all the buccal areas miniscrews can be inserted in certain patients,and the best buccal area is around maxillary second molar roots.Nevertheless,there isn’t an absolutely safe area.The application of dentition distalization by miniscrews in the buccal area of maxillary molar roots needs individual analysis.

Abstract:Objective To explore the relationship between the actual bicarbonate reference value of Chinese women and geographical environment,and provide scientific basis for  the distribution law of  actual bicarbonate of women reference value   and clinical  diagnosis.Methods 8 138 cases of actual bicarbonate reference values were collected.The relationship between reference values of actual bicarbonate of Chinese women and altitude,annual sunshine hours,annual mean air temperature,annual mean relative humidity,annual precipitation,annual range of air temperature were analyzed by correlation analysis.And the geographical factors were as nerve cell input back propagation(BP) neural networks to build nonlinear model.Results The correlation between the reference values of actual bicarbonate of Chinese women and 6 geographical factors was found,and there  were significant differences(P<0.05).The reference value of actual bicarbonate of women in 4 383 points all over China were calculated by a model which was established by 5-layer neural network and 3000 time’s training.The distribution map of the reference values of actual bicarbonate was drawn by spatial interpolation.Conclusion  The reference value of actual bicarbonate of women can be got by artificial neural networks (ANNs) model.The distribution map reflects that the spatial distribution law of   actual bicarbonate reference value in  western is lower than that in eastern of  China.

Abstract:Objective To investigate and calculate the iodine intake of urban residents in different regions in China and assess the results with the criterion in order to offer advice to reasonable iodine intake for people in daily life. Methods Food frequency questionnaire and the 7th weighing methods were used to collect the data of iodine intake of urban residents of three different regions，which were Longyan city of Fujian province,Zibo city of Shandong province and Wuwei city of Gansu province,in China from April 2009 to April 2010. Results The iodine intakes of the urban residents of Fujian Province,Shandong Province,and Gansu Province were 68.03,74.00,and 98.30 μg?d-1,respectively. The iodine contribution rates of iodized salt among four condiments were 99.48%,99.62%,and 98.46%,respectively. Conclusion The iodine intakes of urban residents in three different regions in China are lower than 150 μg?d-1,so the inhabitants still need iodine supplementation from iodized salt.

Abstract:Objective To study the exposure levels of extremely low frequency electromagnetic fields in two workplaces,and to observe the health effects of occupational groups exposed to extremely low frequency electromagnetic radiation in two workplaces.Methods The intensities of electromagnetic fields in two workplaces(control group and exposure group) were detected with EFA-300 frequency electromagnetic field strength tester,and the intensity of the noise was detected with AWA5610D integral sound level.Professional information,household information and personal health condition of 183 controls and 521 subjects in exposure group were investigated by multi-center simple random sampling method.The data was analyzed by SPSS 17.0 statistic software.Results The intensities of electric fields and the magnetic fields in exposure group were significantly higher than those in control group(P<0.05),but the intensities of the noise in two workplaces showed no difference(P>0.05).The questionnaires survey data showed that compared with control group, the incidence of symptoms of loss of hair  in exposure group was significantly higher (P<0.05) and the incidence of decreased hearing was lower （P<0.05）.Conclusion Exposure to extremely low frequency electromagnetic radiation may have some effects on the nervous system of workers.

Abstract:Objective To analyze the epidemiological features of hemorrhagic fever with renal syndrome（HFRS）in Inner Mongolia from  2001 to 2010,where  vaccination-based control strategies were carried out,and to provide a scientific basis for prevention and control of HFRS in the future.Methods The monitoring data of HFRS in Inner Mongolia from 2001 to 2010 was statristically analyzed with Excel software.Results A total of 2 640 HFRS cases and 35 deaths reported in Inner Mongolia with case fatality rate  1.33%,the average annual incidence was 1.10/100 000 from 2001 to 2010.The incidence numbers declined by 34%.The epidemics areas of HFRS had expanded,but there was general sporadic distribution,the HFRS cases mainly concentrated in the Hulunbeier City and Bayannaoer City.The most infections concentrated in the autumn-winter season,the peak was still in November.The cases mainly distributed in the young and adult.The incidence reported in males was more than that in the females at all groups.Farmer had the highest incidence in all occupations.The incidence of the HFRS began to show descending trend year by year,the annual incidence of the HFRS basically  maintained at the low level of less then 1/100 000 after 2007,after three years of implementing expanded program on immunization (EPI),the incidence numbers declined by 543,the average annual incidence declined to 0.43/100 000 from 1.19/100 000,compared with last three years.Conclusion Implement vaccination-based control strategies  had been performed since  2001,and the incidence and mortality of HFRS have greatly decreased in Inner Mongolia.

Abstract:Objective To  establish a rapid accurate method to detect four kinds of enteric pathogens including Listeria monocytogenes,Escherichia coli O157,Proteusbacillus vulgaris,Vibiro parahaemolyticus with multiplex PCR.Methods The special primers were designed of these bacteria for multiplex PCR respectively.Using this screening system to detect the commercially available juice which had been artificially randomly contaminated.Results Multiplex PCR could effectively amplify the corresponding gene fragments,the amlification fragments of Listeria monocytogene,Eschericha coli O157,Proteusbacillus vulgaris and Vibiro parahaemolyticus were 155,366,522 and 199 bp.The sensitivity of the pathogens  reached 103CFU/mＬ;as a result,any of three kinds of bacteria randomly contaminated in the juice could be identified successfully.Conclusion The use of multiplex PCR make it possible that whether the food is carrying the  four pathogenic bacteria mentioned above.