Abstract:Objective　To study the change of c-myc gene expression in thymic lymphomas of mice induced by ionizing radiation and provide theoretical basis for     mechanism of radiation carcinogenesis.Methods　The thymic lymphomas models of BALB/c mice were made by   X-ray(dose rate:0.287 Gy?min-1;total dose:7 Gy) exposuring,6 months later they were killed,then total RNA  and  total protein were extracted  from thymic lymphomas a
nd normal thymus tissue and cDNA was synthesized, the expressions of c-myc mRNA and protein in thymic lymphomas induced by ionizing radiation and normal thymus were detected by quantitative real-time PCR and Western blotting respectively.Results　The expression of c-myc mRNA  in thymic lymphomas（9.16±4.66） was higher than that in normal thymus tissue（1.16±0.54）（P<0.01）. The expression of c-myc protein in thymic lymphomas was higher than that in normal thymus tissue. Conclusion　There is relevance between c-myc gene and thymic lymphomas induced by ionizing radiation，c-myc gene might be susceptibility gene of radiation carcinogenesis.

Abstract:Objective　 To construct the recombinant eukaryotic expression vector of  HMGA2 gene and observe its expression in eukaryotic cells,and laid a theoretical foundation for further study of　HMGA2 gene function.Methods　HMGA2 gene full-length cDNA from human breast epithelial cell HBL-100 was amplified by RT-PCR;after double-digestion and gal recovery HMGA2 gene was cloned into FP-C1 eukaryotic expression vector.After eukaryotic expression vector YFP-C1-HMGA2 was transfected into human prostate cancer cells,and the correctness of constructed YFP-C1-HMGA2 was certificated by RT-PCR.Results　HMGA2 gene full-length cDNA of 351 bp was obtained by RT-PCR.After the cloning of YFP-C1 eukaryotic expression vector,double enzyme digestion and sequencing analysis,YFP-C1-HMGA2 eukaryotic expression vector was confirmed to be successful.And it could express　 in　eukaryotic cells.Conclusion　 HMGA2 gene is cloned successfully,and YFP-C1-HMGA2 eukaryotic　expression vector is constructed successfully.

Abstract:Objective
To investigate the effect of recombinant human basic fibroblast growth factor (rhbFGF) on myocardial angiogenesis in rats with acute myocardial infarction (AMI) and discuss the potential to extend treatment  to coronary artery disease patients who are not optimal candidates for conventional methods by therapeutic angiogenesis.Methods Myocardial infraction was induced in rats by using coronary artery ligation.A total of 60 rats were randomly divided into model group,rhbFGF-H group(50 mg•L-1),rhbFGF-L group (25 mg•L-1) and sham group;rhbFGF was directly injected into ischemic myocardium via some points.Morphometric and histological analysis were used to evaluate infarct size,vascular density and myocardial damage at 24 h,2 weeks and 4 weeks.Results The infarct sizes in the rhbFGF-H group and rhbFGF-L group were decreased significantly and lower than those in model group at 24 h,2 weeks and 4 weeks(P<0.05).The capillary densities in the infarct border zone in rhbFGF-H group and rhbFGF-L group were higher than those in model group and sham group at 2 weeks and 4 weeks（P<0.01,P<0.05）.Hematoxylin-Eosine staining result showed that there were cell swelling,cardial musle fibers breakage,a part of myocyte cytolysis and inflammatory infiltration in the infarct border zone in model group at 24 h,2 weeks and 4 weeks
.Although there were cell swelling in rhbFGF-H group and rhbFGF-L group,but few cardial musle fibers breakage and myocyte cytolysis could be seen. Conclusion rhbFGF can promote the angiogenesis and decrease the infarct size in ischemic myocardium of rats with AMI.

Abstract:Objective
To study the function of AIRE on inducement and function of the CD4+CD25+Treg cells by regulating Notch1 expression,and discuss the mechanism of AIRE in peripheral immune tolerance.Methods  RAW264.7 cells were divided into transfected group and untransfected group,PEGFPC3-AIRE plasmid  was transfected with liposome,RT-PCR was used to detect the expressions of AIRE mRNA and Notch1 mRNA in every group.RAW264.7 cells were divided into transfected group,untransfected group,transfected and anti-Notch1 blocking group,untransfected and anti-Notch1 blocking group;FACS and RT-PCR were used to detect the number of CD4+CD25+Treg cells and the expression of Foxp3  mRNA in every group.Results AIRE gene was successfully transfected into the RAW264.7 cells.Notch1 mRNA expression in transfected group was significantly higher than that in untransfected group.The number of CD4+CD25+Treg cells and Foxp3 mRNA expression in transfected group were significantly higher than that in untransfected group,after applying the Notch1 antibody,compared with control group,the number of CD4+CD25+Treg cells and Foxp3 mRNA expression were lower in experimental group.
Conclusion AIRE could influence the production and function of CD4+CD25+Treg cells by up-regulation of Notch1 for keeping the function on peripheral immune tolerance.

To find the most effective way to obtain highly homogenous and undifferentiated human adipose-derived mesenchymal stem cells（hADSCs）,and induce the hADSCs to differentiate into adipocytes.Methods hADSCs were isolated from human adipose tissue by collagenase digestion and adherence to flasks.hADSCs were differentiated into adipocytes by using dexamethasone,indomethacin,IBMX and insulin.Results hADSCs had unique immunophenotypes and they were positive for CD73,CD44,CD166,CD105 and CD29,but negative for CD31,CD34,CD45 and HLA-DR.hADSCs had the typical proliferative characteristics of stem cells，most of them were in the resting phase ，only a few cells were in the active proliferative phase.They could be induced to differentiate into adipocytes.Conclusion  It is successful to isolate and obtain highly homologous hADSCs,and it was  also successful  in inducing the hADSCs to differentiate into adipocytes in vitro.

To observe the change endoplasmic reticulum stress in renal injury of fluorosis rats, and explore the effect of endoplasmic reticulum stree in the mechanism of renal injury of fluorosis.Methods 48 Wistar rats were divided into 4 groups:control,fluoride-treated,low calcium,and low calcium plus fluoride-treated.The rats in fluoride-treated and low calcium plus fluoride-treated groups were treated with sodium fluoride (NaF,221 mg•L-1) through drinking water for 3 months.The  pathological  slice of rat kidney was made and the  morphological changes were observed under  optical microscope.The transcription levels of Grp78,Xbp1,CHOP and PDI in the  kidney ti
ssues were determined by reverse transcription-polymerase chain reaction (RT-PCR).Results Edema and vacuolar degeneration of the proximal tubule and distal tubule cells in the  kidney tissues in fluoride-treated group were observed;and cell edema
,vacuolar degeneration,scattered necrosis,minor regeneration repair and interstitial hyperemia in the kidney tissues in low calcium plus fluoride-treated group were found
.The mRNA expressions of Xbp1  in the kidney tissues in fluoride-treated  and low calcium plus fluoride-treated groups  were markedly higher than that in   the responding control groups（P<0.05）. The mRNA expression of Grp78     in the kidney tissues in low calcium plus fluoride-treated group  was　significantly increased   than that in  fluoride-treated and  low calcium groups（P<0.05 or  P<0.01）.The mRN
A expression of PDI   in the  kidney tissues in low calcium plus fluoride-treated group was significantly decreased than that  in  fluoride-treated group （P<0.05）;there was no significant difference in CHOP expression in the kidney tissues among 4 groups.Conclusion Overdosal fluoride can induce renal injury,and low calcium nutrition may  aggravate the toxicity of fluoride on kidney.Endoplasmic reticulum stress is likely involved in the pathogenesis of renal injury of fluorosis rats.

 To investigate the effects of small interference RNA (siRNA) targeting vascular endothelial growth factor (VEGF) on VEGF expression and cell proliferation of cervical carcinoma HeLa cells,in order to provide a theoretical basis for the treatment of human cervical carcinoma.
Methods siRNAs plasmids targeting VEGF165 were constructed and stably transfected into HeLa cells.HeLa group,Mock group and VEGF siRNA group were set up.The expressions of VEGF mRNA and protein in stable transfectants were confirmed by RT-PCR,Western blotting and ELISA assay,respectively.The cell proliferation was analyzed by MTT and flow cytometry.Apoptosis was measured by Hoechst33342 staining.Results VEGF siRNA stably transfected cell lines were successfully established.The siRNA targeting human VEGF gene effectively decreased the expression of VEGF at RNA and protein levels,and inhibited the proliferation of HeLa cells (P<0.05).The inhibitory rates of cell proliferation in  HeLa group,Mock group and VEGF siRNA group at 24,48 and 72 h were 12.8%±5.4%，17.4%±3.7%, and 27.2%±5.7%.Compared with Mock group, VEGF siRNA significantly inhibited the cell proliferation in HeLa cells(P<0.05).Hoechst33342 staining result showed that VEGF siRNA did not have obvious effect on the apoptotic rate of HeLa cells  compared with Mock group.
Conclusion  The siRNA targeting human VEGF gene could effectively reduce the expression of
 VEGF both at RNA and protein levels,and inhibit the  proliferation of  cervical carcinoma cells in vitro.It shows that VEGF plays a role in the development of cervical carcinoma and it may be beneficial in finding new gene therapy for cervical carcinoma.

To explore the influence of tumor cells in the number and function of CD4+CD25+Treg cells.
Methods  Lewis lung cancer cells and mouse spleen lymphocytes co-culture system was established.Lewis lung cancer cells with different concentrations of lymphocytes co-cultured were divided into 4 groups:experimental group Ⅰ(5×105 Lewis lung cancer cells and 1×106 lymphocytes c
o-culture),control group Ⅰ(1×106 lymphocytes culture),experimental group Ⅱ(5×105 Lewis lung cancer cells and 2×106 lymphocytes co-culture),control group Ⅱ(2×106 lymphocytes culture);Lewis lung cancer cells were co-cultivated with lymphocytes for different time, 24,48, and 72 h three time points were selected.Lewis lung cancer cells culture supernatant was co-cultivated with lymphocytes,the concentrations of culture supernatant were 20% and 50%.The number changes of CD4+CD25+Treg cells in the co-culture system of Lewis lung cancer cells and splenic lymphocytes were detected by flow cytometry;the expression of Foxp3 mRNA after co-culture was detected by RT-PCR method.
Results Compared with control group,the number of CD4+CD25+Treg cells and the expression of Fo
xp3 mRNA were significantly increased in experimental group Ⅰ (P<0.05),and ther
e was no significant difference in experimental group Ⅱ (P>0.05);24　and 48 h 　
after co-culture of Lewis lung cancer cells and lymphocytes,the number of CD4
+CD25+Treg cells and the expression of Foxp3 mRNA were significantly increased (P<0.05),
and there was no significant changes at  72 h;20% and 50% Lewis lung cancer cells supernatant could significantly increase the number of CD4+CD25+Treg cells and Foxp3 mRNA expression (P<0.05).Conclusion Tumor cells and their supernatants could induce the increase of the number of CD4+CD25+Treg cells  and their function,this might be one of mechanisms of tumor-induced immune tolerance.

To detect the inhibitory effects of ionizing radiation combined with autophagy and apoptosis inhibitors and inducers on the proliferation of human breast cancer cell line.
Methods  MTT and flow cytometry (FCM) were used to detect the surviving and proliferation of MCF-7 cells，which were under 0,2,4,8 and 10 Gy X-ray radiation and different dealing methods 4 Gy,4 Gy + 3-MA,4 Gy + rapamycin,4 Gy + z-VAD-fmk,and   the relationship of dose-effects and time-effects was analyzed.
Results  With the increase of irradiation doses (4,8 and 10 Gy) and the elongation of irradiation time (48 and 72 h),the inhibitory rates of the  proliferation of breast cancer cells were increased ,there were significant differences between various groups(P<0.05 or P<0.01).The inhibitory rates of the proliferation of breast cancer cells in  4 Gy+3-MA or 4 Gy+ z-VAD-fmk groups  were significantly different from those in 4Gy+rapamycin group(P<0.05 or P<0.01),and there were significant differences after treated for  24,48 and 72 h between various groups (P<0.05 or P<0.01).Conclusion Ionizing radiation in combination with autophagy inducer could induced the autophagy in human breast cancer cells and promote the apoptosis;the ionizing radiation in combination with autophagy inhibitor or apoptosis inhibitor could inhibit the apoptosis.Thus,ionizing  radiation can induce the autophagy in human breast cancer cells,and promote the apoptosis.

Abstract:Objective To explore the effect of modified amantadine(NAM) on immune function of the mice infected with influenza A(H5N1) virus and to provide experimental basis for development of NAM.Methods Mouse models infected with influenza A (H5N1) virus were established.And the mice were divided into 7 groups:normal control group（NC）,positive control group (PC),amantadine group (AMA) and 4 NAM groups (which contained 25,50,100 and 200 mg?kg-1 NAM, respectively).The spleen and thymus index,T lymphocyte transformation,NK cytotoxicity activity and interferon (IFN) activity of mice were detected.Results The  mean spleen index in 100 mg?kg-1 NAM group was higher than that in PC group (P<0.05),and the mean thymus index in NAM group was higher than that in PC group (P<0.05).The spleen indexes  in 25,100 and 200 mg?kg-1 NAM groups were increased compared with PC group(P<0.05),and were and increased in 100 and 200 mg?kg-1 NAM groups compared with AMA group (P<0.05).The NK cytotoxicity activities in 100 and 200 mg?kg-1 NAM groups were   higher than that in PC group(P<0.05),while they were also higher in 25,100 and 200 mg?kg-1 NAM groups than that in AMA group (P<0.05).The IFN activities in 50,100 and 200 mg?kg-1NAM groups were increased compared with PC group(P<0.05),and the IFN activities in 50,100 and 200 mg?kg-1 NAM groups were also increased compared with AMA group (P<0.05).
Conclusion NAM can stimulate the proliferation of spleen and thymus cells of mice infected with influenza A(H5N1) virus,and enhance T lymphocyte transformation,NK cytotoxicity  and IFN activity.

Abstract:Objective
 To explore the reparative effect of bactericidal/permeability-increasing protein(BPI) on the damaged mucosa of rats with  otitis media with effusion (OME)，and state the pathogenesis of OME.
Methods  Wistar rats(40 ears) were randomly divided into six groups:normal control group (n=4),BPI control group(n=4),eustachian tube obstruction (ETO) group (n=8
),lipopolysaccharide (LPS) injection group (n=8), ETO+LPS group (n=8)，ETO+LPS+BPI group (n=8).The experimental OME model was made through eustachian tube obstruction and LPS injection.The rats were killed after 1,2 and 4 weeks and the changes of mucosa of middle ear were observed under light and scanning electron microscope.
Results  The rats in normal control group and BPI control group had the normal mucosa in the tympanic orifice of the eustachian tube.It consisted of pseudostratified ciliated cubical or columnar epithelium which contained an abundant number of ciliated cells and a few goblet cells,these were the mucociliary clearance system of the middle ear.The hypotympanum consisted of thin,squamous epithelium with few microvillus.Middle ear mucosa was obviouly thickened in LPS injection,ETO and  ETO+LPS  groups.An increase in goblet cells and a decrease in ciliated cells were observed in the tympanic orifice of the eustachian tube.The epithelial layer in the hypotympanum had become more pseudostratified ciliated cubical epithelium.In ETO+LPS+BPI group,there was thin squamous epithelium in the hypotympanum near normal,which was not thickened and contained few microvillus.
Conclusion  LPS and  ETO can result in  the occurrence and protracted courses of OME by mimosa’s inflammatory reaction which can reduce the activity of ciliary cells and weaken the  function of mucociliary clearance system.BPI could bind avidly to LPS,reduce inflammatory reaction,and break the inflammatory cycle and reestablish an effective mucocillary clearance system.The  results suggest that  BPI treatment is a potential effective drug  for prevention   and therapy of  chronic OME.

 To investigate the changes of phosphorylated CREB in spinal cord dorsal horn of rats underwent chronic compression of dorsal root ganglia (CCD) and effects of intrathecally administer NOS inhibitor L-NAME,AG or 7-NI and cGMP  analogue 8-Br-cGMP  on the expression of pCREB in dorsal horn and the changes in thermal hyperalgesia of rats.Methods 90 male adult Wistar rats were randomly divided into L-NAME,AG,8-Br-cGMP,7-NI,Cremophor and PBS groups.Different NOS inhibitors were injected intrathecally by microsyringe at the 5th day after CCD,the thermal paw withdrawal latencies were measured before injection and  0.5,2.0,6.0,12.0,24.0 h after treatment,the anti-nociceptive effects of agents were compared.The expression of pCREB was detected by immunohistochemical analysis 2 h after intrathecal injection.Results CCD  significantly increased the expression of pCREB and pCREB positive neurons located in  all laminae of bilateral spinal cord.The expression of pCREB in ispilateral and contralateral spinal cord showed no significant difference.Compared with PBS  group,there was significant decreasing in number of pCREB positive neurons in L-NAME,AG and 7-NI groups(P<0.01），and there was markedly increasing in 8-Br-cGMP group（P<0.05）;Simultaneously,the intrathecal administer L-NAME,AG and 7-NI could  significantly reverse thermal hyperalgesia induced by CCD
.Conclusion  NO-cGMP-PKG signal pathway contribute to CCD-induced neuropathic pain.

To investigate the effects of purine nucleotides compensation on the withdrawal symptoms，contents of 5-hydroxytryptamine（5-HT），5-hydroxyindoleaceticacid （5-HIAA）and their  potential mechanisims.Methods 
 Rats were administered with heroin by intraperitoneal injection with increasing dose to develop addiction models．One hundred and ten male Wistar rats were divided into five groups：control group (C)，heroin group (H)，heroin plus purine mononucleotides (AMP+GMP，equal mole mixture) group (HAG)，heroin plus adenosine monophosphate (AMP) group (HA)，heroin plus guanosine monophosphate (GMP) group (HG).The contents of 5-HT and 5-HIAA were analyzed by fluorospectrophotometry.The content of tryptophan hydroxylase (TPH) was detected by ELISA. The expression of TPH protein was detected by immnunohistochemistry.Eight  rats from each group were used for the observation of acute withdrawal symptoms induced by naloxone at 4 h after drug administration on the 10 th day, four  rats from each group were used for immunohistochemistry.
Results  Withdrawal symptoms were alleviated in HAG，HA and HG groups (P<0.05). The contents of 5-HT and 5-HIAA in H group were significantly decreased compared with C group (P<0.05).While in HAG，HA and HG groups，the contents of 5-HT and 5-HIAA were increased when compared with H group (P<0.05).The TPH content was decreased in H group when compared with C group (P<0.05)； in HAG，HA and HG groups，the TPH contents were increased when compared with H group (P<0.05)；the expression of TPH protein was decreased in H group when compared with C group (P<0.05)，and it was increased in HAG，HA and HG groups when compared with H group (P<0.05).
Conclusion  Purine nucleotides can partly alleviate withdrawal symptoms and increase the contents of 5-HT and TPH in heroin dependent rats.

Abstract:Objective To investigate the effects of polyi:c on angiogenesis in mouse prostate carcinoma tissue and their possible mechanisms.Methods Mouse prostate carcinoma models were randomly divided into two groups according to tumor weight:control group and polyi:c group.After  treatment for 7 times,the mice were sacrificed and the tumor tissues were cut for weighing,calculating the tumor inhibitory rate and tumor index.Hematoxylin-eosin staining and immunohistochemical staining were performed to observate the morphological changes of prostate carcinoma tissues,distribution of vasa and expressions of vascular endothelial growth factor (VEGF) and endothelial nitric-oxide synthase (eNOS).
Results In polyi:c group,the mean tumor inhibitory rate was 67.85% and the tumor index was（5.42±0.17）%;in control group,the tumor index was（14.45±1.06）%;  there was significant differences between polyi:c group and control group( P<0.01). The expressions of VEGF in polyi:c group and control group were (9.64±2.90)% and (37.91±7.62)%, respectively,and there was significant difference between two groups( P<0.01). The expressions of eNOS in polyi:c group and control group were (4.43±10.39)% and (22.00±8.07)%，respectively,and there was significant difference between two groups ( P<0.01).Conclusion Through decreasing the expressions of VEGF
and eNOS,polyi:c plays an important role in inhibitting the angiogenesis and indirect inhibiting mouse prostate carcinoma.

Abstract:Objective To explore the medicinal values of the buds of Lonicera ruprechtiana Regel.by analyzing its chemical constituents. Methods The buds of Lonicera ruprechtiana Regel.were extracted with 70% ethanol,the extractives were purified with  macroporous resin and isolated with silica gel and ODS column chromatography.NMR,UV,IR Spectrometers were used to obtain spectroscopic data.
Results According to the single spot in TLC,five compounds were isolated from the buds of Lonicera ruprechtiana Regel.,and their structures were identifited through physico-chemical constants and spectroscopic analysis.Five compounds were isolated from the buds of Lonicera ruprechtiana Regel.,and they are daucosterol (Ⅰ);7S-O-methylmorroniside(Ⅱ);neohesperidin(Ⅲ);benzyl-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside (Ⅳ);6′-O-β-D-apiofuranosyl sweroside (Ⅴ).Conclusion  All these compounds are isolated from the buds of Lonicera ruprechtiana Regel.for the first time.

Objective
 To explore the mechanism of ginsenoside-Rh2(G-Rh2) on inhibition of glioma by identifying differential proteins with proteomic technique. Methods   The total proteins were extracted from SHG-44 cells treated with 32 μmol?L-1 G-Rh2 for 72 h and the cells in control group,then were subjected to two-dimensional gel electrophoresis.Only spots with a fold change equal or above 1.5 and P<0.05 were selected  as differential proteins.Afterwards the differential proteins were analyzed by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for peptide mass fingerprint (PMF) identification. 
Results  Compared with those of control group,20 differential protein spots were identified by the two-dimensional electrophoresis in SHG-44 cells treated with G-Rh2,including 16 down-regulated ones and 4 up-regulated ones.Five remarkably down-regulated proteins analyzed by mass spectrometry were cofilin 1,phosphoglycerate kinase (PGK),peroxiredoxin 1 (Prx 1),heat shock proteins (HSP) and proliferation cell nuclear antigen (PCNA). Conclusion  These differential proteins may be involved in the proliferation inhibition of human glioma cells by G-Rh2.

Objective To investigate the inhibitory effect of hCAP-18/LL-37 gene transfection on proliferation in Lewis cells,and to study the possible antitumor mechanism.Methods DNA of hCAP-18/LL-37 transient transfection mediated by Polyfect was employed.The experimental cells were classified into untransfected cells,pcDNA4/Myc-His transfected cells and pcDNA4/Myc-His- LL-37 transfected cells.The proliferating abilities of Lewis cells 48 and 72h after transfection were measured by MTT colorimetric assay.The apoptosis of transfected Lewis cells was detected by flow cytometry and  fluorescence microscope.The expressions of Bax and Bcl-2 in transfected Lewis cells were determined by cellular and immunohistochemical technique.Results The proliferating viability of Lewis cells transfected by the recombinant plasmid was significantly inhibited,the inhibitory  rate of growth  was 3.51% and 17.65% compared with pcDNA4/Myc-His transfected cells.The apoptotic rate were 7.4%,4.4%,5.2% in untransfected cells,pcDNA4/Myc-His cells and pcDNA4/Myc-His- LL-37 cells, respectivly, by FCM measurement.The protein expression of Bax in transfected Lewis cells was increased,and there was significant difference compared with control(P< 0.05).The protein expression of Bcl-2 in transfected Lewis cells was decreased,but there was no significant difference compared with control(P>0.05).
Conclusion hCAP-18/LL-37 gene transfection can induce apoptosis of Lewis cells by  up-regulation of Bax and  down-regulation of Bcl-2 and inhibit the proliferating ability of Lewis cells.

Objective To detect the expressions of Bcl-2 and Bax gene and protein  in myocardium of rats with isoproterenol (Isp)-induced myocardial necrosis and significances.Methods 40 Wistar rats were divided into control group,12 h,1 week and 3 weeks  after Isp injection groups.Except control group,all the rats in the other groups were treated with isoproterenol hydrochloride.12 h,1 week and 3 weeks after Isp injection,immunohistochemistry and RT-PCR were applied to detect the gene and protein expressions of Bcl-2 and Bax.Results The results of immunohistochemistry showed that  12 h and 1 week after Isp injection,the expressions of Bax were increased significantly compared with control group (P<0.05);Inversely,the expressions of Bcl-2 were decreased significantly 12 h,1 week and 3 weeks after Isp injection compared with normal control group (P<0.05).The ratios of Bcl-2/Bax were significantly decreased  12 h,1 week and 3 weeks after Isp injectioncompared with control group (P<0.05). The expressions of Bax mRNA were increased significantly
 12 h,1 week and 3 weeks after Isp injection compared with control group (P<0.05);The expressions of Bcl-2 were decreased significantly 12 h and 1 week after Isp injection compared with control group (P<0.05).Conclusion Apoptosis of myocardium is the basic pathological change of myocardial fibrosis,and the expressions of gene and protein of Bcl-2 family  play an important role in this process.

Objective
To investigate the expressions of FSH receptor mRNA and protein in ovary tissue in rats with  letrozole-induced polycystic ovary syndrome(PCOS),and to provide experimental data for the model application. Methods Forty rats were randomly divided into two groups(n=20),in PCOS model group letrozole  was administered once daily during 21 d,and  in control group  without any treatment.The  gonadal hormone concentrations in serum were determined by radioimmunoassay,the histologic changes in ovaries were observed by HE staining,the expression of FSH receptor gene in ovary tissue was detected by realtime -PCR,Western blotting and immunohistochemistry.
Results  Compared with control group,estradiol(E2) and progesterone in model group showed a considerable reduction（P<0.05）,the serum testosterone (T) and follicl
e-stimulating hormone (FSH) and luteinizing hormone(LH) levels in model group were markedly increased than those in control group（P<0.05）.The ovarian weights of the two groups had no significant difference(P>0.05).Compared with control group,the ovaries from model group showed high incidence of subcapsular ovarian cyst and capsular thickening and decreased number of corpora lutea.The expressions of FSH receptor mRNA and protein were significantly higher in model group than those in control group（P<0.05）.Conclusion The expression of FSH receptor gene in letrozole-induced polycystic ovaries is similar with that of PCOS women,the rat model is proved to be an ideal PCOS animal model to study the pathophysiology of PCOS.

Objective To investigate the effects of octreotide  (OCT) on gene and protein expressions of collagen Ⅰ，collagen Ⅲ and  matrix matrixmetlloproteinases 2 (MMP-2)in hepatic stellate cells（HSC） in vitro，and to clarify the mechanism of its anti-hepatic fibrosis.Methods  A rat hepatic stellate cell line,rHSC-99,was treated with OCT.Cell proliferation was assessed by MTT colorimetric assay.The contents of  collagen Ⅰ，Ⅲ were examined by ELISA；RT-PCR was used to detect mRNA expressions of MMP-2,collagen Ⅰ and collagen Ⅲ.Results Compared with control group,OCT significantly inhibited rHSC-99 proliferation in vitro (P<0.05),the inhibitiory rate of cell proliferation was increased with the elevation of the OCT in a certain dose range (1-8 μg?L-1).Compared with control group,the concentrations of collagen  Ⅰ,Ⅲ in OCT groups were reduced  significantly,the difference was significant (P<0.05);and the  mRNA expressions of collagen Ⅰ and Ⅲ  were also decreased (P<0.05).Compared with control group,MMP-2 mRNA was highly expressed in OCT groups (P<0.05).Conclusion OCT could inhibit the proliferation of HSC,down-regulate the expressions of collagen Ⅰ and  Ⅲ mRNA and protein，and up-regulate the expression of MMP-2 mRNA.All of these biological effects of OCT on HSC are associated with recovery from liver fibrosis.

Objective
To construct the recombinant plasmid that highly expressed h
uman subcellular PTEN,inorder to provide a basis for further study on its anti-tumor effect.
Methods PTEN cDNA was amplified by RT-PCR based on mRNA of placenta.The PCR product was ligated into T-vector,and transfected into E.coli;the obtained T-PTEN plasmid was identified with restrictive digestion and sequencing.PCR was  used to incorporte nuclear signal of localization(NSL) into PTEN when T-PTEN was used as template.Then the PCR product  was  ligated into T-vector,and transfected into E.coli,and T-NSL-PTEN plasmid was obtained.pcDNA3.1 and T-NSL-PTEN were ligated after digested with EcoRⅠand BamHⅠ,and transfected into E.coli,the recombinant vector pcDNA3.1-NSL-PTEN was obtained,and identified with digestion and sequcncing.Results The recombinant  expression vector DUM-PTEN and PUM-NSL PTEN were  identified by restrictive digestion and DNA sequencing.As expected,by EcoRⅠ and BamHⅠ digestion,it showed the band of 1 200 bp.The sequencing result showed the NSL was incorporated successfully.The recombinant pcDNA3.1-PTEN was obtained with 1 200 bp,the sequencing result showed that its sequence was same as target gene;the recombinant pcDNA3.1-NSL-PTEN was comfirmed by restrictive digestion and sequencing,and the NSL was incorporated successfully. Conclusion The  recombinant expression plasmid pcDNA3.1-NSL-PTEN is constructed successfully which can highly express human subcellular PTEN.

Objective
To explore the lead-expelling effect of Quqian Yizhi Health Oral Liquid (QYHOL) in lead-poisoning rats,and provide experimental basis for prevention and therapy of lead poisoning. Methods 72 SD rats were randomly divided into six groups. One group was negative control group only gavaging with distilled water,and others were used to establish lead poisoning models by gavaging 2.0% lead acetate  for three weeks.Positive control group was gavaged with distilled water,EDTA group was treated with 0.5% EDTA by abdominal cavity injection and the last three groups were gavaged with low,medium and high-dose QYHOL(1.25,2.50 and 5.00 mg?kg-1 respectively) for three weeks.At the end of the experiment,all rats were killed and the samples of blood,livers,kidneys and femurs were collected.The lead level in each sample was measured by hydride generated-atomic absorption spectrographic method.The pathological  changes of tissue sections of liver,kidney and other organs stained with HE were observed by light microscope.Results Compared with positive control group,the lead levels of rat blood samples,livers,kidneys and femurs were markedly  decreased after treated with QYHOL with three dosages.Among three dosage QYHOL groups,there were no significant differences in the decreasing   percentage of  lead levels in blood and kidney tissues (P>0.05).The decreasing  percentage of lead levels  in liver tissues in low-dosage group was higher than those in high-dosage and EDTA groups(P<0.05 or P<0.01).High-dosage QYHOL markedly decreased the lead levels in femur tissues,and compared with low-,mid-dosage and EDTA groups,there were significant differences(P< 0.01).All pathological sections with HE staining did not reveal abnormal structures by light microscope.Conclusion  QYHOL has safty and effective role on lead-poisoning rats.
         
            

Objective To study the effects of formaldehyde on the level of microelements in mouse liver and blood and explore the microelements as biomarker in injury induced by formaldehyde.Methods The mice were randomly divided into control,low（1/40LD50）,moderate（1/20LD50） and high（1/10LD50） dose groups exposed to formaldehyde by inhalation(n=20),two hours per day for three and six weeks,and than the contents of Cu,Fe,Zn and Mn in mouse liver and blood were measured.Results  The contents of Fe,Zn and Mn in liver in high dose group three weeks after exposure and the contents of Fe and Zn in liver in high dose group six weeks after exposure were significantly lower than those in control group(P<0.05);the contents of Mn and Zn in blood in high dose group three weeks after exposure were significantly lower than those in control group(P<0.05);while six weeks after exposure,the contents of Fe and Zn in blood in moderate and high dose groups were significantly lower than those in control group(P<0.05);and the content of Cu had no significant change in each group.
Conclusion Formaldehyde might decrease the contents of Fe,Zn and Mn in mouse liver and blood,and the Fe,Zn and Mn could be used as biomarkers in injury induced by formaldehyde.

Objective
 To determine the existence of RBM6-RBM5 chimera and study its relationship with oncogenesis. Methods Ten cell lines including GLC-20,MDA.MB.231, Jurkat,TF-1,MCF-7,Hela,BT-474,A431,K562 and Raji ； several cDNAs from tumor tissues including lung,ovary,skeletal,skin,spleen,uterus,heart,bone,brain,lymph node,pancreas,prostate,testis  and their normal controls and 5 breast cancer tissues were collected for the study.Nested PCR was used to identify the existence and expression level of RBM6-RBM5 chimera.QPCR was used to determine the RBM6,RBM5 gene expression level in chimeric positive and negative tumor tissues.Results RBM6-RBM5 chimera was identified in MDA，MB，231 cell line,which lacked exons 20,21 of RBM6 and exon 1 of RBM5.The chimeric expression was only observed on 10 tumor cell lines and tissues,among some specific tumor types,the chimeric expression was associated with elevated levels of RBM6 and RBM5 mRNA.Conclusion  RBM6 mRNA experiences altered co-transcriptional gene regulation in certain cancers.RBM6-RBM5 transcription-induced chimerism might be a process that is linked to the tumor-associated increased transcriptional activity of the RBM6 and RBM5 genes,and this chimera might be a new marker for tumor prediction. 
〖WTHZ〗Key words：RNA binding motif protein;transcription-induced chimera;tumor cells,cultured

Objective  To explore the antioxidant effect of honeysuckle    on hepatic RBL cells of rats in vivo  and in vitro and its underlying mechanism,and  provide theoretical basis for the application of honeysuckle in clinic.Methods The blood levels of T-AOC,SOD,GSH-Px,GSH or MDA in an antioxidant enzyme system were detected 1-2 h after administration in the established rat models,which were administrated with larger and smaller doses of honeysuckle respectively.The cultured human hepatic RBL cells were exposed to H2O2 as an oxidant and treated with 0.25g?L-1 neysuckle
before or after exposure to H2O2.The cell survival and apoptosis were detected by MTT and TUNEL assays.The expression levels of NF-kB,Bcl-2,Bax,HSP-70 and  Caspase-3 were determined by immunohistochmistry and Western blotting and the cell anti-oxidant enzyme system was detected by corresponding kits.Results In the antioxidant and anti-apoptotic effects of honeysuckle,the pre-protective effect was markedly higher than the post-protective.The expressions of NF-κB and HSP-70,the expression ratio of Bcl-2/Bax were down-regulated,and the apoptotic rate in  pr
e-protective group was lower than that in  post-protective group.  Besides,the detected cell level of anti-oxidant enzyme system was up-regulated significantly.In the plasma of rat model the levels of T-AOC,GSH-Px,GSH and SOD were promoted(P<0.05),while that of MDA was decreased markedly(P<0.05).Conclusion  Honeysuckle displays antioxidant capacity through up-regulating the antioxidant enzyme system and down-regulating NF-κB signal transduction pathway.

Objective
 To determine the protective effect of extracorporeal circulation with autologous lung as oxygenator to lung function in dogs.Methods Twelve adult mongrel dogs were randomly divided into control group and experimental group.Cardiopulmonary bypass (CPB) using a membrane oxygenator (control group) or using the autologous lung (experimental group) for gas exchange was performed for 120 min in an alternating series of 12 mongrel dogs with the heart arrested for 90 min by crystalloid cardioplegia and 30 min reperfusion.Malondialdehyde (MDA) of lung tissue,alveolar-arterial oxygen gradient (P［A-a］O2),the ratio of white blood cells in BALf and lung function were measured at different time points （before CPB, 15 and 60 min after CPB）,lung biopsies were also made. Results The levels of airway resistance at  15 and 60 min after CPB in control group were higher than the airway resistance levels before CPB (P<0.05).The levels of airway resistance at  15 and 60 min after CPB in experimental group  were lower than those in control group at the same points (P<0.01).The levels of pulmonary vascular resistance at 15 and 60 min after CPB in each group were lower than the levels before CPB (P<0.01).The level at  15 and 60 min after CPB in control group  was higher than that  in experimental group  at the same points(P<0.05).The levels of PaO2/FiO2 at  15 and 60 min after CPB in each group were higher than the levels before CPB (P<0.01).The levels of PaO2/FiO2 at  15 and 60 min after CPB in control group   were lower than those  in experimental  group at the same points(P<0.05).The levels of P［A-a］O2 at  60 min after CPB in experimental group  were lower than those  in control group(P<0.001).The levels of MDA and neutrophil percentage in BALF at  90 min after CPB in  experimental group were lower than those 
 in control group (P<0.001,P<0.05).Morphological analysis revealed that  in  experimental group the  percentage of pathological lesions and alveolar hemorrhage grade values was lower than those in control group (P<0.001).
Conclusion Morphous and function have obvious changes in lungs after CPB.Extracorporeal circulation with autologous lung as oxygenator has  protective effect on  pulmonary morphous and function.

Objective
To study the effect of Tongbiding freezing-dryied  powder  for injection on the function of WDR neuron of spinal cord in living rats with chronic constriction injury (CCI), and  clarify its analgesia mechanism in  spinal cord  center, and further explore   whether it has the drug dependence at  the cellular level. Methods Electricity physiology extracellular recording technique was used , in the CCI living  rat’[KG-*3]s expanded spinal cord waist stage  the electricity activity of identical WDR neuron cells was recorded  before and after administration of 20 mg.kg-1 Tongbiding. The electric discharge number of C response was observed before  and  2, 4, 8, 10 min after administration. The spontaneous electric dischare number. was observed  before and   1, 2, 4, 6 min after administration; wind-up phenomenon was also observed.Results The electric discharge number of C response  was obtained
 in  8 WDR neurons,three were significant differences of the mean electric discharge number of C response  between 2,4,8 min after administration and before administration (P<0.05 or P<0.01). The observation of cell’[KG-*3]s spontaneous ele
ctric discharge number recorded  12 cells altogether, 6 cells stopped discharging spontaneously immediately after administration, 6 cells stopped discharging spontaneously  6 min after administration. There were significant differences of the mean  spontaneous electric discharge number  between 2, 4 min after administration and before administration  (P<0.01).Five  wind-up phenomena were found altogether, this phenomenon was suppressed after .
 Conclusion The analgesia effect of Tongbiding freezing-dryied   powder  for injection on spinal cord  center  is related to affecting the WDR neuron, and it is confirmed  that the drug does not have drug dependence  at  the cellular level.

Objective
To study the effects and mechanism of Yunnan Baiyao on the inflammatory response and satellite cell regeneration after acute injury of skeletal muscle in rats. Methods
Rat models of skeletal muscle injury (gastrocnemius, fast twich;soleus,slow twich) induced by treadmill were constructed 24 h after establishment of the models,  the left sides of hind limbs in rats were treated with   Yunnan Baiyao, and the right sides were treated with saline at the equal volume.4 rats were chosen 2,4,7,10,14 d after injury,respectively,and  gastrocnemius and soleus of the treated sides (the left sides) and untreated sides (the right sides) from hind limbs of 4 rats were obtained,and stained with  immunohistochemistry and HE. The quantity   and morphological changes  of inflammatory cells and satellite cells  were observed under optical microscope. Results Compared with untreated sides, the number of inflammatory cells of fast twich after injury in  the treated sides was  decreased obviously at the  2nd,4th,7th day after injury(P<0.01) and the number of inflammatory cells of slow twich  at the 2nd day  after injury in treated sides was  decreased (P<0.01);the  necrotic cells of skeletal muscle were decreased.Compared with untreated sides  the muscle  fibers of fast twich  and slow twich  were repaired at the 7th,10th day,and the number of satellits cells was increased obviously in the treated sides(P<0.01). Conclusion Yunnan Baiyao can inhibit the destruction of inflammatory cells to injuried  muscle tissues, shorten the inflammatory reaction process and promote the proliferation of satellite cells and muscle fiber repairment after injury.

Objective
To investigate the genetic association between the polymorphism of rs12026099 site,rs2250312 site and rs1022528 site on PTGER3 gene and schizophrenia in Han population from the north of China.Methods PCR-RFLP analysis was employed to detect the genotype of PTGER3 gene in 240 family trios consisting of fathers,mothers and affected offsprings with schizophrenia.Haplotype relative risk (HRR) analysis,transmission disequilibrium test (TDT),Haplotype analysis and association between SNPs and clinical symptoms analysis were used to analysis the genotype data.
Results The genotypic frequency of the PTGER3 gene did not deviate from Hardy-Weinberg equilibrium in both patient group and parent group (P>0.05);HRR analysis did not show allelic association with the PTGER3 gene (χ2= 1.396,P>0.05;χ2= 0.042,P>0.05;χ2= 0.119,P>0.05);TDT analysis did not show preferential transmission of the two alleles (χ2= 1.380,P>0.05;χ2= 0.043,P>0.05;χ2= 0.126,P>0.05);Haplotype anal
ysis showed that the two SNPs were in one haplotype block,but the haplotype was not associated with schizophrenia(χ2=6.500,P>0.05);χ2 test for association betwee
n SNPs and clinical symptoms of schizophrenia showed that rs12026099 locus was associated with apathy of schizophrenia and illogic thought of paranoid type schizophrenia (χ2=4.578,P=0.032;χ2=6.100,P=0.047),rs2250312 locus was associated with illogic thought and abulia of paranoid type schizophrenia (χ2=4.471,P=0.034;χ2=4.206,P=0.040),rs1022528 locus was associated with illogic thought of paranoid type schizophrenia (χ2=4.108,P=0.043). Conclusion rs12026099 locus,rs2250312 locus and rs1022528 locus on PTGER3 gene are not associated with schizophrenia,but associated with clinical symptoms of schizophrenia in Han population from the north of China.

Objective
To investigate the susceptibility to essential hypertension(EH) in Ningxia Hui population based on the distribution of   the polymorphisms of HLA-DQB1 in  EH population . Methods   PCR-SSP method was conducted to examine the genotypes of HLA-DQB1 in 66 healthy individuals and 84 EH patients,and the genotypic and allelic frequencies were statistically computed. Results The distribution of alleles was significantly different between two groups.The frequencies of *0301,*0401,*0601,*0604/5/6/7,*0302 in EH group were significantly higher than those in control group (P<0.01 or P<0.05).The frequencies of *0501,*0602,*0603/8,*0606,*0402,*0604/5/6/8 in EH group were significantly lower than those in control group (P<0.01 or P<0.05).Conclusion In Ningxia Hui population,allele DQB1*0301,*0401,*0601,*0604/5/6/7 and *0302 might be danger factors, while *0501,*0602,*0603/8,*0606,*0402 and *0604/5/6/8 might be resistive factors of EH.

Objective
 To investigate the changes of Th1/Th2 cytokines levels in serum in patients with  chronic hepatitis B patients befroe and after treatment of thymosin,and clarify the relationship with state of illness,curative effect and virological response.
Methods 19  patients with chronic active hepatitis B were chosen as treatment group, 10 cases as normal control group.The patients in  treatment group were treated with  injection of thymosin α 1 mL,in the pre-treatment,13 and 21 weeks after treatment the cytokine levels,HBV DNA,and biochemical indicators of liver function were detected by flow cytomertry on Tuesday,Friday every week;in normal control group the  serum IL-2,IFN,TNF,IL-4,IL-6,IL-10 indicators were determined and the relationship between  cytokine levels and biochemical responses and  virological response was analyzed.Results  After thymosin treatment,the cytokine levels secreted by Th cells were increased compared with  pre-treatment (P<0.05), the levels of Th1 cytokines (IL-2,IFN,TNF) in reponse group were higher than those in non-response group, the levels of Th2 cytokines (IL-4,IL-6,IL-10) in non-response group were higher than  those in response group (P< 0.05).According to  biochemical response  analysis,the cytokine levels in patients could reflect the curative effect, the cytokine levels in  response group were lower than those in partial response group,the cytokine levels in  part  response group were lower than those in non-response group,the cytokine levels in response group  were lower than those in non-response group,the increase of cytokine levels had negative correlation with curative effect of virological response  (P<0.05).
Conclusion The changes of the levels of serum Thl/Th2 cytokines  in patients with  chronic hepatitis B after treatment of thymosin may  contribute to the removal of  HBVDNA in body.

Objective
To discuss the effects of carvedilol on the heart function and glucolipometabolism in elder patients with diastolic heart failure.Methods 76 elder patients with diastolic heart failure were randomly divided into treatment group(n=34) and control group（n=42）,all the patients were given routine treatment of diastolic heart failure,the patients in treatment group were given carvediol in addition( from little dose to more ),the course of treatment was 3-6 months.The parameters of the diastolic function of heart before and after treatment in two groups were determined by ultrasonic cardiogram and uninjured cardiodynamic machine.And the levels of fasting blood glucose,triacylglucerol(TG),cholesterol in two groups were detected.
Results After treatment the total effective rate in treatment group was higher than that in control group( P<0.05).There was no significant difference of blood glucose and lipid between before and after treatment in treatment group.But compared with control group,the levels of blood glucose and lipid in treatment groupo were decreased(P<0.05).During the treatment there was no significant adverse effects in two groups,no stopping treatment.Conclusion Carvedilol could improve the function of left ventricle of elder patients with diastolic heart failure,and has no significant effects on the metabolism of glucose and lipid.

Objective
To investigate the changes of homocysteic acid (Hcy) and C-reactive protein (CRP) levels in blood plasma of elderly patients with coronary heart disease and elucidate the risk factors of elderly patients with coronary heart disease.Methods 90 elderly patients with coronary heart disease were divided into 3 groups:stable angina pectoris group (SAP)(n=30),unstable angina pectoris group(UAP)(n=30),acute myocardial infarction group(AMI)(n=30).30 healthy elderly persons elder than 60 years old were enrolled as control group.Venous blood of all persons was harvested during empty stomach and plasma was separated.Then plasma Hcy and CRP were determined.
Results The TC and TG levels in the three elderly coronary heart disease groups were significantly higher than those in control group （P<0.05）. Especially the TG level increased following with the increase of severe degree of elderly coronary heart disease.The Hcy and CRP levels in the three elderly coronary heart disease groups were significantly higher than those in control group （P<0.05）.There were significant differences of  plasm Hcy and CRP levels between three elderly coronary heart disease groups（P<0.05）.The plasma Hcy and CRP levels increased following with the increase of severe degree of elderly coronary heart disease.Conclusion Elderly coronary heart disease is related to Hcy emia and the increase of CRP level.This demonstrates that Hcy emia and the increase of CRP level may be risk factors for elderly coronary heart disease.At the same time,the abnormality of blood fat is related to Hcy  emia and the increase of CRP level.Following with the increase of severe degree of elderly coronary heart disease,the plasm Hcy and CRP levels increase,too.So severe degree of elderly coronary heart disease can be predicted through determination of plasm Hcy and CRP levels.

Objective
 To study the expressions of survivin and caspase-9 in human primary hepatocellular carcinoma(HCC)　 tissues and their relationship with apoptosis.Methods Immunohistochemical staining was uesed to detect the expressions of survivin and caspase-9  in paraffin sections of 96 cases including 42 HCC tissues,42 paired adjacent noncancerous tissues and 12 normal liver tissues. Apoptosis was detected by TUNEL.Results No immunoreactivity of survivin was seen in normal liver tissues.The positive rates of survivin in HCC and paired adjacent noncancerous tissues were 73.81% and 4.76%,respectively,there was significant difference between two groups(P<0.01). The positive rates of caspase-9 in normal tissues,paired adjacent noncancerous tissues and HCC tissues were 83.33%,80.95% and 47.62%, respectively,and the positive rate of caspase-9 in HCC tissues was significantly lower than those in the other two kinds of tissues (P<0.05),but the expression of caspase-9  in paired adjacent noncancerous tissues had no difference compared with normal tissues (P>0.05).The expressions of survivin and caspase-9 in HCC tissues were not associated with age,sex and the size of tumor,but they were directly associated with tumor histological grade and metastasis.Conclusion  High expression of survivin in HCC may inhibit apoptosis through depressing the expression of caspase-9,and it is an indicator of independent poor prognosis.

Objective
To investigate the effect of low level laser （LLL）irradiation with different doses on proliferation and differentiation of human osteoblast-like cells,and provide the experimental basis for clinical application of LLL.Methods Human osteoblast-like cells MG63 were irradiated by Ga-Al-As diode laser (808 nm,66 mW) with doses of 1 J?cm-2 and 3 J?cm-2 after being seeded into multi-well plates.Untreated cells were used as controls.The proliferation of MG63 was quantified by colorimetric MTT (dimethylthiazol tetrazolium bromide) assay.The expressions of the marker of differentiation osteopontin （OPN） and collagen Ⅰ （Coll Ⅰ） were observed by immunohistochemistry assay.Results There was no difference of proliferation and expressions of OPN and Coll Ⅰ 　between the laser group with dose of 1 J?cm-2 and control group(P>0.05). The laser group with the dose of 3 J?cm-2 showed more increase in cell survival at 24 and 48 h after being seeded as compared with control group(P<0.05).Moreover,the expressions of OPN and Coll Ⅰ in the laser group with dose of 3 J?cm-2 was also higher than those in control group (P<0.05).
Conclusion LLL irradiation with the dose of 3 J?cm-2 may promote the proliferation and differentiation of human osteoblast-like cells,which might improve bone healing.

Abstract:Objective To identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells，and investigate the relationship between them and  select cervical cancer stem cell markers.Methods  Flow cytometry was used to identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells；after isolation,the expressions of CK17 and CD44v5 in Bcrp1+ and Bcrp1-  phenotype HeLa cells were measured by  immunocytochemistry.Results HeLa cells contained 11% Bcrp1+ cells,0.7% CK17+ cells and 0.1%CD44v5+ cells.Bcrp1+ HeLa cells contained CK17+ and CD44v5+ HeLa cells,but the Bcrp1- Hela cells did not（P<0.05）.Conclusion HeLa cells expressed CK17 and CD44v5 antibody must express Bcrp1 antibody.CK17 and CD44v5 may be used as the markers of cervical cancer stem cells.

Objective
To study the expressions of CD24 and SLeX in lung cancer tissues,and  evaluate the relation ship with  development and progression of lung cancer.Methods Immunohistochemistry was used to detect the expressions of CD24 and SLeX in 70 specimens of lung cancer.Results The positive expression rates of CD24,SLeX in non-small cell lung cancer tissues were 41.3% and 63.5% respectively，There were no expression and weak expression respectively in small cell lung cancer (SCLC) tissues. The positive rates of CD24,SLeX in adenocarcinoma were  55.9% and 76.5%,respectively,which were higher than those in squamous cell carcinoma (24.1%,48.3%)(P<0.05).In lung adenocarcinoma,aquamous cell carcinoma,there was no correlation between CD24-positive patients with age,gender,smoking index,tumor stage and grade(P>0.05).In lung adenocarcinoma and squamous cell carcinoma,the positive expression rate of SLeX  in stage Ⅲ (83.3%) was higher than that in stage Ⅰ-Ⅱ (55.6%),the positive rate in poor differentiated lung cancer(80.8%) was higher than those in  moderate and well differentiate cancer(51.4%),the positive rate in female patients (76.7%) was higher than that in male patients (51.5%)(all P<0.05).The average survival time of the patients with positive expressions of SLeX and CD24 was significantly lower than that of the patients with negative expressions (P<0.05).Conclusion CD24 and SLeX can  play advance roles in the occurrence and development of lung　cancer ， and they can be used as indicators to judge prognosis in lung cancer.

Objective
 To study the expressions of progesterone receptor A (PRA) and B (PRB) in breast cancer and adjacent non-malignant tissues and the correlations between their  expressions and the clinical characteristics. Methods The expressions of PRA and PRB in 50 specimens of female human breast cancer and adjacent non-malignant tissues were detected by immunohistochemistry. The correlations between the expressions of PRA and PRB and the clinical characteristics were analyzed.Results PRA and PRB expressed in both the nuclei and the cytoplasma of tumor cells and epithelial cells of the acini and ducts.The percentages of  PRA and PRB positive cells were 42%,42% and 52%,36% in the cancer and the adjacent non-malignant tissues, respectively,there was no significant difference.The expression of PRA was significantly correlated with the age of patient(r＝－0.316 8，P<0.05) in adjacent non-malignant tissues.The expressions of PRA and PRB were significantly correlated with the expression of CDC-47（r＝－0.422 8，P<0.05；r＝－0.529 5，P<0.05）and ERα(r＝0.403 2，P<0.05；r＝0.378　6，P<0.05) in malignant tissues.The expressions of PRA in malignant tissue or  adjacent non-malignant tissues were significantly correlated with the expression of PRB (r＝0.840 1，P<0.05；r＝0.424 6，P<0.05). Conclusion PRA and PRB have synergetic effect,but PRA may play a more important role in the development and progression of breast cancer.

Objective
To study the expressions of MIF,CD68 and CD57 which are the markers of macrophages,macrophage migration inhibition factors and natural killer cells in ovarian cancer tissues and their significances. Methods Immunohistochemistry staining was used to detect the expressions of MIF,CD68 and CD57 in 56 ovarian cancer tissues and 5 normal ovary tissues. Results  MIF,CD68 and CD57 had positive expressions in ovarian cancer tissues at different degrees,but the expressions of MIF,CD68 and CD57 were week or negative in normal ovary tissues.Furthermore,the positive expression levels of MIF,CD68 and CD57 in ovarian cancer tissues were increased with the grade of ovarian cancer.The expression of CD57 was lower than the expression of CD68 in ovarian cancer（P<0.05）.Conclusion The up-regulated expression of MIF can make the increase of macrophages and natural killer cells,it is one of the reasons of tumor development. 

Objective
 To examine the effects of weight loss on functional state of female judokas. Methods  Twelve female judokas belonging to Shanghai female judo team were tested at different periods(first,second,third phases and last phase) during weight loss a month before competition. The changes of physiological and biochemical indexes in different periods were observed and all the data  was  statistically analyzed. Results  Previous three phases, the  Fat%  in weight loss group was decreased compared with basic data (P<0.05); at the last phase, the lean body weight(LBW),back power and grip power were decreased compared with basic data and control group (P<0.05). Compared with basic data and control group,the hemoglobin (Hb) and RBC in weight loss group were decreased, then the hematocrit(Hct) was increased (P<0.05). Compared with basic data, the blood urea nitrogen(BUN)  in weight loss group  was decreased at the third phase (P<0.05). At the last phase, BUN and cortisol (C) in weight loss group  were increased, then testosterone(T) was decreased compared with basic data and control group (P<0.05).Conclusion Weight loss before competition has the effects on physiological and biochemical indexes, which lead to the decline of functional state and exercise ability of female judokas.

Objective
To study the clinical effect of Qiliqiangxin capsule (QC)  in patients with  chronic congestive heart failure (CHF). Methods According to random number table,64 CHF patients were randomly assigned into therapy group (n=33) and control group (n=31).During the treatment,sweat secretion,heart function improved rate,6 min walking distance,the side-effect and compliance were observed.The levels of serum Na+ and K+ of patients were measured on the 2nd,8th and 15th hospital day.Results  Compared with control group,the remission rate  of sweat secretion in therapy group was higher after treatment (P<0.05).One week later,heart function and 6 min walking distance were improved in both groups,but there was no significant difference between two groups（P>0.05）;compared with control group,the improved rates of heart function and 6 min walking distance in therapy group were higher after two weeks (P<0.05).There was no significant difference between two groups of the level of serum Na+ on the 8th hospital day（P>0.05）；on the 15th day,the level of serum Na+ in therapy group was higher than that in control group (P<0.05). There were no significant differences between two groups of the level of serum K+ on the 8th or 15th hospital day（P>0.05）.During treatment,33 patients had not any uncomfortable complain about this drug and nobody stopped using this capsule;while there was no complain from the patients of control group.Conclusion  The effect of QC about releasing hyperidrosis in a short time is conspicuous.QC can improve heart function,but it plays this role gradually.QC can keep the level of serum Na+ to some extent.There is no influence on the level of serum K+.The tolerance of this capsule is well.

Objective
To study the value of clinical application of 1H proton magnetic resonance spectroscopy(1H MRS) in the differential diagnosis of intracranial lesions with ring-like enhancement. Methods 28  cases were diagnosed of intracranial lesions with ring-like enhancement by clinical examination and pathologic test.A total of 28 ratios cases included  6 cases high grade glioma, 10 cases of metastatic carcinoma(n=10) and 12 cases of brain abscess,after examined with 1HMRS,the ratios of various metabolites in focal center,enhancement ring,perifocal edema region and normal control group were detected and compared.Results The ratios of NAA/Cho,Cho/Cr and NAA/Cr in focal center had no significantly differences between high grade glioma and metastatic carcinoma（P>0.05）. The peak of NAA was significantly different between high grade glioma and metastatic carcinoma（P<0.05）. The ratios of NAA/Cho and Cho/Cr in the perifocal regions of high grade glioma and metastatic carcinoma were significantly different compared with normal brain (P<0.05). The relative content of Cho was not significantly different between perifocal regions of metastatic carcinoma and normal brain（P>0.05）. The peak of AA was characteristic of brain abscess. The ratio of Cho/Cr0  in brain abscess was significantly lower than those in high grade glioma and metastatic carcinoma（P<0.05） (Cr0 denoted the Cho content of contralateral normal brain region).These resutls  accorded with the result of pathological examination. Conclusion  1HMRS can improve the diagnostic accuracy of intracranial lesions with ring-like enhancement.

Objective
To probe the value of clinical application of three dimensional contrast enhanced magnetic resonance angiography (3D CE MRA) for diagnosis of peripheral artery occlusive diseases (PAOD) and to assess its accuracy. Methods One hundred and three cases of PAOD received 3D CE MRA before operation. And 579 vascular segments were displayed. The diagnosis of 3D CE MRA before operation were compared with the results of vascular reconstruction surgeries of lower extremities. The image quality and how 3D CE MRA revealed the abnormal blood vessels were evaluated. Results Satisfactory images of the main arteries of the lower extremities were achieved by 3D CE MRA. In 579 vascular segments of 103 patients with PAOD, the sensitivity and specificity for detection of severe stenosis and occlusions were 97.6% and 95.4%， respectively. In 206 vascular segments of tibiofibula artery, the sensitivity was 100%. In 196 vascular segments of femoral artery, the specificity was 97%. Conclusion 3D CE MRA is accurate and reliable for determination of the degree of peripheral artery stenosis. The results of 3D CE MRA are concordant with that of operation.

Objective
 To determine whether the use of 18F-FDG PET/CT alters staging and management of nasopharyngeal carcinoma (NPC) when compared with MRI staging practice；and to explore the relation of standard uptake value (SUV) of 18F-FDG PET/CT and the pathological classification and T staging of NPC. Methods The study was performed retrospectively on a group of 41 patients with a new diagnosis of NPC. All the patients underwent whole body PET/CT scanning and head & neck MRI scanning within 3 weeks of each other. The AJCC protocol was introduced to stage NPC and the results of the PET/CT were compared with MRI based on pathologic diagnosis. Results ①Primary tumor: the accuracy of T staging of PET/CT was significantly higher than MRI (85.37% vs 60.98%, U=2.49，P< 0.05). According to the pathologic results, PET/CT downstaged 3 patients and upstaged 7 patients. In the patients with NPC, the SUV had  positive correlation with T staging (rs=0.706，P<0.05); the SUV in the patients with undifferentiated carcinoma was significantly higher than that in the patients with poorly differentiated squamous carcinoma (t=7.89，P<0.05). ②Lymph nodes metastases: among all the 41 cases, 139 lymph nodes were obtained. PET/CT correctly detected 72 positive lymph nodes and 56 negative lymph nodes, the sensitivity and specificity were 93.51% and 90.32%,respectively; while MRI correctly detected 60 positive lymph nodes and 51 negative lymph nodes, the sensitivity and specificity were 80.00% and 79.69%,respectively. The accuracy of N staging of PET/CT was significantly higher than MRI(92.09% vs 79.86%,U=2.93，P< 0.05) ③Distant metastases：Among the 41 patients, 2 patients with liver and bone metastases were detected by PET/CT, which missed by MRI. Conclusion  18F-FDG PET/CT stages NPC more accurately and sensitively than does MRI. 

Objective
To explore the application value of  lung volumes from 64 multi-slice CT (MSCT) images in patients with chronic obstructive pulmonary disease (COPD) and study the  correlation between lung volumes measured by MSCT and pulmonary function test (PFT) results. Methods 24 patients clinically diagnosed with COPD (COPD group) and 22 healthy people (control group) were selected and underwent both chest MSCT scans and PFT within one week. The total lung was scanned at full inspiration and full expiration with MSCT, respectively. The total lung volumes were measured by CT Pulmo software (Siemens, Forchheim, Germany). The quantitative total lung volumes from MSCT images were compared with PFT and SPSS13.0 was applied to assess the correlation.  Results Compared with  control group, the full inspiration volume (Vin) (P<0.05), the full expiration volume (Vex) (P<0.05),and Vex/Vin (P<0.01)were elevated, while Vin－Vex was declined obviously in COPD group (P<0.05).Vin was in positive correlation with total lung capacity (TLC) (r= 0.923 ,P<0.01) and Vex was also in positive correlation
with residual volume (RV)(r= 0.912 ,P<0.05), as well as Vin－Vex with vital capacity (VC) (r=0.763，P<0.01) and Vex/Vin with RV/TLC (r= 0.754 ,P<0.01). The Vex showed best correlation with FEV1% and FEV1/FVC (r=－0.616,P<0.01;r=－0.543，P<0.05). Vin,Vex and Vex/Vin were significantly elevated in patients with COPD  compared with those in control group. Conclusion Lung volumes obtained from MSCT images show a strong correlation with PFT results in patients with COPD. Lung volumes measured from MSCT images can be used to evaluate the pulmonary function in patients with COPD.

Objective
To construct single PCR assay for brucella, and optimize reaction conditions through simulative blood samples and inoculated mice, and provide laboratory evidence for PCR assay application in clinical diagnosis of brucellosis following-up. Methods Primer BP26 on genus level according to BP26 gene and three primers on strain level according to IS711  were designed; simulative samples of serial concentrations were  constructed  and mice were inoculated with M5 or Pps858-MCS2/M5 as experimental group, while PBS as control; brucella DNA was extracted by boiling method. This PCR assay was used to detect bacteria solution with primers BP26 and three strain primers,simulative samples  and  inoculated  mice  were diagnosed with primer BP26 and strain primer Meli. Results  Designed primers on genus and strain level showed good specificity through brucella standard strains and vaccine strains;  DNA was extracted from simulative samples and blood samples of the inoculated mice by the boiling method and  specific bands were obtained; the PCR positive numbers of the inoculated mice increaseed early and desceased later, and increasing sample quantities  increased the  positive rate; the PCR positive rate of the boiling method was  higher than chemical method at the same time point, and the PCR positive rate was  more sensitive than traditional culture method. Conclusion The boiling method is convenient to extract DNA from blood. The PCR assay demonstrates higher sensitivity than traditional culture method; this PCR assay is very specific and available to be used for following-up of brucellosis patients.