To study the changes of mRNA and protein expressions of Kras gene in thymic lymphomas induced by ionizing radiation,and to detect the methylation of CpG islands in promoter region of Kras gene,then to investigate the mechanisms for the occurrence of radiation carcinogenesis.Methods The thymic lymphoma models of BALB/c mice were made by X-ray irradiation,then the total RNA was extracted,cDNA was synthesized and the total protein was extracted from both thymic lymphoma tissue and normal thymus tissue;the mRNA and protein expressions of Kras gene in thymic lymphoma tissue and normal thymus tissue were detected by RT-PCR and Western blotting method,and the methylation of CpG islands in promoter region of Kras gene was detected by bisulfite sequencing PCR.Results The mRNA expression level of Kras gene in thymic lymphoma tissue was significantly higher than that in normal thymus tissue（P<0.01）.The protein expression level in thymic lymphoma tissue was about 1.41 times higher than that in normal thymus tissue;4 CpG sites were methylated detected by bisulfite sequencing PCR in normal thymus tissue,however,1 CpG site was methylated in thymic lymphoma tissue,the CpG islands in promoter region of Kras gene were demethylation state in thymic lymphoma.Conclusion Ionizing radiation can cause the changes of mRNA and protein expression levels  of Kras gene in thymic lymphoma tissue   by demethylation state of Kras gene,eventually lead to the occurrence of tumor;it might be one of the mechanisms for the occurrence of radiation carcinogenesis.

Abstract:Objective
To explore the mechanism of radio-resistance of breast cancer stem cells by investigating the effects of ionzing  radiation on the reactive oxygen species (ROS),apoptosis and cycle distribution.Methods The breast cancer MCF-7 cells  were suspension cultured in serum-free medium containing a variety of  growth factors.There were MCF-7 (breast cancer cells),MCF-7-S (breast cancer stem cells),MCF-7+8 Gy and MCF-7-S+8 Gy groups in the experiment.4-24 h after 8 Gy irradiation,the ROS levels,percentages of apoptotic cells  and percentages of the cells at each cycle phage   were measured by FCM with 2′,7′-dichlorodihydrofluorescin diacetate (DCFH-DA),AnnexinⅤ-FITC/PI and PI staining,respectively.Results The breast cancer stem cell microsphere accumulated hundreds of cells were obtained successfully at 7 d after suspension culture with serum-free medium containing a variety of growth factors;the FCM results showed that CD44+CD24－ phenotype breast stem cells were up to 75.20%.With the time prolongation,the ROS levels and apoptosis in MCF-7 group and MCF-7-S group showed increasing trendency,and reached for the maximum values at 12 and 24 h;the ROS levels in MCF-7-S group were significantly lower than those in MCF-7 group at 4,8,12 and 24 h (P<0.05 or P<0.01),and the   percentage of apoptotic cells in MCF-7-S group was significantly higher than that in MCF-7 group only at 8 h (P<0.05);the ROS levels (4,8,12 and 24 h) and   percentage  of apoptotic cells(12 h) were significantly increased in MCF-7+8 Gy group (P<0.05),and the  percentages of apoptotic cells (4,8,12 and 24 h) in MCF-7-S +8 Gy group were significantly decreased (P<0.05 or P<0.01),but the ROS levels had no obvious change in MCF-7-S+8 Gy.At 12 h,as compared with MCF-7 group,the  percentages of the cells at  G0/G1 phase and G2/M phase in MCF-7-S group were significantly decreased (P<0.05),and the  percentage of the cells at S phase was significantly increased (P<0.05 );the  percentage of the cells at G2/M phase in MCF-7+8 Gy group was significantly increased (P<0.05 ),but there were no significant changes in MCF-7-S+8 Gy group.Conclusion Ionizing irradiation can cause the increasing of ROS level and apoptosis and G2/M phase arrest in breast cancer cells,but has no obvious effects on the breast cancer stem cells;it indicates that radio-resistance might be related to ROS level,apoptosis and G2/M phase arrest.

Abstract:Objective  To research the relationship between miR-21 and radiation sensitivity of cervical cancer HeLa cells,and to elucidate the possible action mechanism of miR-21 in apoptosis and autophagy of cells.Methods The HeLa cells were divided into mock groups (transfected with miR-21 NC,mimics, inhibitor NC or inhibitor) and 4 Gy irradiation groups(transfected with miR-21 NC,mimics, inhibitor NC or inhibitor with irradiation).The miR-21 expression was detected by real-time PCR;colony-forming assay was used to detect the changes of radiation sensitivity;the apoptosis and autophagy were analyzed by FACS assay;the target gene of miR-21 was predicted by bioinformatics methods;Dual-luciferase Reporter Assay was used to demonstrate the binding of miR-21 and targets;the expression of beclin1 protein was measured by Western blotting method.Results After 4 Gy radiation,compared with mock group,the expression of miR-21 was increased remarkably (P<0.05). Colony-forming assay showed that there were no changes of radiation sensitivity of HeLa cells after overexpressing or inhibiting miR-21  compared with miR-21 NC or miR-21 inhibitor NC groups(P>0.05). Compared with miR-21 NC＋4 Gy group,the apoptotic rate of HeLa cells in　miR-21 mimics+4 Gy　group was  increased (P<0.05).After transfection with  miR-21 inhibitor,compared with miR-21 inhibitor NC  group,the apoptotic rate of HeLa cells in duced by radiation was  decreased (P<0.05).Compared with miR-21 NC  group,the autophagy rate was increased after transfection with miR-21 mimics (P<0.05).After transfection with miR-21 mimics and radiation,the expression level of beclin1 protein was increased(P<0.05) compared with miR-21 NC+4 Gy group.Conclusion Beclin1 is the target gene of miR-21 in HeLa cells.Overexpression of miR-21 can increase the radiation-induced apoptosis and autophagy,but has on influence in radiation sensitivity of HeLa cells.

Abstract:Objective
To analyze the influence of dexamethasone in the hypotonic-induced volume-sensitive chloride currents in human trabecular meshwork cells,and to investigate the possible mechanism of volume-sensitive chloride channels(VACC) in the glucocorticoid-induced glaucoma(GIG) cases.Methods The human trabecular meshwork cells were seeded in 35 mm diameter plastic petri dishes,so that they could grow in monolayer.The cultured cells were divided into  normal cell culture medium group and dexamethasone 1 d,3 d,7 d groups.The  chlorine current density values of the cells in four groups were recorded respectively by the whole-cell patch-clamp technique.The differences among groups were compared.Results In normal group,after hypotonic stimulation,under +100 and -100 mV voltage clamp,the outward and inward current density values of the trabecular cells were (19.94±0.87) and (－6.53±0.41)　pA/pF.In dexamethasone 1 d,3 d， and 7　d groups,under  the same condition,the outward and inward current density values of the trabecular cells were  (19.39±1.40) and (－6.42±0.28)　pA/pF,(17.97±2.35) and (－5.82±0.94)　pA/pF,(17.16±1.16) and (－5.65±0.43)　pA/pF.The trabecular cells cultured with dexamethasone for 1 d  had lower outward and inward current density values under hypotonic stimulation compared with normal group,but there was no significant difference (P>0.05).The trabecular cells cultured with dexamethasone for 3 d and 7 d,when compared with normal group,had significantly lower outward and inward current density values under hypotonic stimulation (P<0.05).Conclusion Dexamethasone could reduce the volume-sensitive chloride current in trabecular meshwork cells,which would affect trabecular meshwork cell volume adjustment.This would possibly cause the increase of the aqueous humor outflow resistance among GIG cases.

Abstract:Objective To construct an eukaryotic expression vector of human B-cell translocation gene 2 (BTG2),to  express the FLAG-tagged BTG2 protein in HeLa cells,and to supply an experimental tool for investigating the function of BTG2 gene.Methods The full-length BTG2 fragment was obtained by PCR and inserted into the multiple cloning site of pcDNA3.1(+) vector.Oligo DNA encoding FLAG tag was designed and inserted into pcDNA3.1(+)-BTG2 to construct another vector pcDNA3.1(+)-FLAG-BTG2.The HeLa cells were divided into pcDNA3.1(+) empty vector group,pcDNA3.1(+)-BTG2 group and pcDNA3.1(+)-FLAG-BTG2 group.The HeLa cells were transfected with recombinant plasmids.Western blotting using anti-FLAG antibody was performed to detect the expression of FLAG-BTG2 protein in HeLa cells.Results The sequence of the vector was verified by both BamH Ⅰ endonuclese digestion and DNA sequencing.The Western blotting analysis confirmed that FLAG-fused BTG2 was detected in pcDNA3.1(+)-FLAG-BTG2 group but not in empty vector or pcDNA3.1(+)-BTG2 groups. Conclusion The eukaryotic expression vector pcDNA3.1(+)-FLAG-BTG2 is successfully constructed and FLAG-tagged BTG2 protein is expressed in HeLa cells.

Abstract:Objective  To set up the molecular cytobiological model of  endogenous coagulation factor Ⅷ (FⅧ) re-expressing in human liver cells L02, and to study the regulation pathway and molecular basis of the re-expression of FⅧ in L02 cells activated by NO signal.Methods The L02 cells at logarithm growth phase were selected and randomly divided into blank control group and experimental group,inhibitor group and  inhibitor control group;they were cultured for 0,12,24,36,48, and 60 h.Flow cytometry was used to detect the expression of human FⅧ protein in L02 cells after treated for 48 h.Griess experiment was performed  to detect the levels of NO in L02 cells at different time points;the transcription levels of human FⅧ gene,iNOS gene,NF-κB1 gene and I-κB alpha gene were detected by RT-PCR method.Western blotting method was used to detect the expression levels of human phosphorylated I-kappaB (phosphorylated I-κB)  in L02 cells.Results The results of flow cytometry showed that the expression of human L02 FⅧ protein was found after treated with L-arginine for 48 h.The Griess results showed that the levels of NO in L02 cells in experimental group were significantly increased at 3,6,12, and 24 h（P<0.05） and the levels of NO in blank control group,inhibitor group and  inhibitor control group had no changes.The RT-PCR results showed that the transcription of human FⅧ mRNA in L02 cells was found in experimental group,but there was no transcription of human FⅧ mRNA in blank control group,inhibitor group and  inhibitor control group;the transcription levels of iNOS,NF-κB1 and I-κB alphain experiment group were increased（P<0.05） and the transcription levels of these genes in blank control group,inhibitor group and  inhibitor control group had no changes.The Western blotting results showed that after adding L-arginine the expression level of phosphorylated I-κB was significantly increased（P<0.05）,other groups had no such change.Conclusion L-arginine can activate the phosphorylation of I-κB by NO signal pathway to lead to the changes in the expression of human FⅧ gene promoter upstream regulatory-related transcription factors NF-κB to activate the expression of human FⅧ in human liver cells L02.

Abstract:Objective  To study the  apoptosis of K562 cells induced by a new immunosuppressive agent FTY720 and its mechanism,and to provide experimental basis for the treatment of leukemia in clinic.Methods The K562 cells were cultured in vitro　and  divided into blank control  group and FTY720 treatment  group.The K562 cells in FTY720 treatment  group were treated with 6 μmol?L-1 FTY720  for 3,6,and 12 h,or treated with different concentrations of FTY720 (2，4，6,8 μmol•L^-1 ) for 24 h.The apoptosis,level  of reactive oxygen species(ROS), mitochondrial membrane potential(MMP) and cell cycle were measured by flow cytometry.The  inhibitory rate of proliferation of  K562 cells after treated with FTY720 was detected by WST-1 reducting assay.Results The results of  flow cytometry showed that the percentages of apoptotic  cells were increased  after treated with 6 μmol•L^-1    FTY720 for 3,6, and 12 h with the prolongation of time compared with blank control group（P<0.01）.The percentages of apoptotic cells were also increased  after treated with different concentrations of FTY720 for 24 h compared with blank control group（P<0.01）. Compared with blank control group,the ROS levels were  increased with the increasing of  FTY720 concentration,while the MMP was decreased（P<0.01）.Compared with blank control group,the percentage of cells at G0／G1 phase was increased,while those at S and G2/M phases were decreased with the increasing of  FTY720 concentration (P<0.05).The WST-1 reduction assay results indicated that the  inhibitory rates of proliferation of K562 cells after treated with 2,4,6, and 8 μmol•L^-1  FTY720 for 72 h were (24.0±4.1)%，(46.4±3.9)%，(67.0±4.8)%, and (88.2±5.6)%, respectively,compared with blank control group.The concentration of FTY720 to result the   inhibitory rate of 50% (IC50) on K562 cells was 5.5 μmol•L^-1 .Conclusion FTY720 could inhibit the proliferation of K562 cells   by blocking  cell cycle and  inducing  apoptosis through provoking ROS.

Abstract:Objective
To observe the growth characteristics and multi-time dynamic contrast-enhanced MR(DCE-MR) findings of rabbit VX2 liver tumor on MR imaging，and to provide a reliable basis for the values of MR in assessment of non-surgical treatment effect of rabbit VX2 liver tumor.Methods 10 rabbit VX2 liver tumor models were established by liver tissue embedding through laparotomy under direct vision  and DCE-MRI scaning was performed at the 2nd,3rd,4th and 5th week after establishment.The tumor volume (V) and tumor growth rate（TGR)  were calculated at different time points and the signal characteristics were analyzed on MR imaging;then all the rabbits were excuted and   HE staining were performed to observe the cancer cells in different places with microscope.Results The TGR at the  3rd week after establishment （404.16%±114.64%） was significantly higher than that at the  4th week （223.49%±65.90%）（t=3.417，P<0.05）.The tumors with well-boundary were rim-like enhanced significantly and the central parts were not enhanced on DCE-MR imaging,and the parts between the edge and centre of tumors were heterogeneously mild enhanced.The HE staining results showed that 6 cases of tumor were squamous cell carcinoma and the cells were distributed densely in the edge,the cancer cells and necrosis were distributed equally in the parts between the edge and centre of tumors and the centre of tumors were necrosis parts.Conclusion The rabbit VX2 liver tumor grows rapidly at the 3rd week after establishment,and its histological features can be better reflected by DCE-MRI.

Abstract:Objective
To research the jugular foramen,internal auditory pore (IAP) and the turning point between its components by imageological methods and to provide theoretical basis for retrosigmoid approach in the operation of acoustic neurinoma.Methods The skulls of 100 volunteers were scanned to get the final result with thin-section  computed  tomographic  image.High-resolution spiral  CT  multiplane reformation was used to reform images that were parallel to the Frankfort horizontal plane to measure the distance between the turning point of retrosigmoid (A),the edge of jugular foramen(B) and the lower edge of the internal auditory canal(C)　(denoted AC,AB,BC) and the shortest distance from the jugular foramen to AC.The angles between AC,AB and sagittal axis(α,β) were measured.Results The distance of AC was  （44.94±3.84） mm,the distance of AB was (43.68±4.56) mm.The distance of BC was （6.15 ±2.04） mm,and the shortest distance between jugular foramen and AC was  （5.21±0.23） mm.The angle α was measured as （39.50±4.74）°,and the angle β was measured as （46.35±5.51）°.Conclusion The research measure the distance and angle between entry points and landmarks of retrosigmoid approach and the safe distance.

Abstract:Objective
To acquire some related data of surgical approach through brain superior temporal sulcus to temporal horn of lateral ventricle by MRI volume rendering,and to orientate the point of superior temporal sulcus on the lateral surface which is closest to temporal horn of lateral ventricle,and to find out the best entrance point of surgical approach through superior temporal sulcus to temporal horn of lateral ventricle.Methods 120 adult cases of MRI scanning specimens were chosen for measurement.MRI volume rendering technology was used to rebuild the brain 3D model for the measurement of the full length of superior temporal sulcus S1.Then cutting along the prependicular to the direction of the long axis of the temporal lobe with 1.0 mm spacings,the coronal sections were obtained,and the distance from superior temporal sulcus to temporal horn of lateral ventricle was ordinally  measured and the shortest distance S2 was made sure.And the depth of superior temporal sulcus S3 was detected.The corresponding point on the surface of the brain at superior temporal sulcus according to the point leading the shortest distance S4 was determined.The ratio of S4 to S1 M was calculated.The angle between the shortest distance and median sagittal plane as α was determined.All the samples were measured on both sides of the brain and all the data were compared.Results The S1 of the 120 cases was (159.56±17.55) mm on the left and (164.35±15.07) mm on the right，there was no statistical difference between two cerebral hemispheres(P>0.05）;the S2 was (8.18±0.96) mm on the left and (7.81±0.90) mm on the right,there was no statistical difference between two cerebral hemispheres(P>0.05）;the S3 was (12.19±1.43) mm on the left and (11.57±1.33) mm on the right,there was no statistical difference between two cerebral hemispheres(P>0.05）;the S4 was (100.88±16.09) mm on the left and (104.15±14.49) mm on the right,there was no statistical difference between two cerebral hemispheres(P>0.05）;the M was (0.63 ±0.07) on the left and (0.63 ±0.06) on the right,there was no statistical difference between two cerebral hemispheres(P>0.05）;the α was (55.80±3.64)° on the left and (56.46±4.17)° on the right,there was no statistical difference between two cerebral hemispheres(P>0.05）.Conclusion The point at the front side 3/5 of superior temporal sulcus may be the ideal surgical approach entrance point.The distance from the point to temporal horn of lateral ventricle is shortest.It indicates that the approach can reduce the damage  of brain tissue.

Abstract:Objective To measure the reconstructed cranial CT images,and to clarify the safety range of unilateral nasal transsphenoidal approach for pituitary surgery.Methods 100 normal pituitary cranial CT images were randomly selected,and the three-dimensional reconstruction was performed by using the CT images of perpendicular and parallel to the edge of the two eyes as base line,and the distance and angle in unilateral nasal transsphenoidal approach for pituitary surgery from the sagittal plane in the middle of the nasal meatus and the plane through the tip of the nose and both ends of dorsum sellae were measured, respectively.The angles and distances were compared when grouped the data by gender and age.Results Angle A1(the angle between the tip of the nose and the tuberculum sellae and saddle back root line in the sagittal plane) in the sagittal plane of the middle nasal meatus was（11.22±1.35）°,95% confidence interval was 8.92°-13.76° degrees; the distance D1(the distance on the line between tuberculum sellae and saddle back root,and the line was formed by the plate contained the angle A1 and sellar floor) was （16.71±2.07） mm,95% confidence interval was 13.11-19.93 mm.Angle A2(the angle between the tip of the nose and the saddle back ends) which was in the plane through the tip of the nose and both ends of dorsum sellae was （8.91±1.19）°,95% confidence interval was 7.12 °-10.72°; the distance D2（the distance on the line between the saddle back ends，and the line was formed by the plate contained the angle A2 and sellar floor) was （14.23±2.09）mm,95% confidence interval was 10.81-17.92 mm. The four parameter data was normally distributed,and there was no significantly statistic difference between different gender and ages (P> 0.05).Conclusion The angle of the movement for unilateral nasal transsphenoidal approach for pituitary surgery operation in the sagittal plane in the middle of the nasal meatus should be less than (11.22±1.35) °,and the distance of the movement should be less than (16.71±2.07) mm. The angle of the movement in the plane through the tip of the nose and both ends of dorsum sellae should be less than （8.91±1.19）°,and the distance of the movement should be less than （14.23±2.09） mm.

Abstract:Objective To explore the effect of different arch forms of maxillary models on stress distribution of peri-implant of All-on-4 implants with 3-dimensional finite element,and to provide evidence for the improvement of implant achievement ratio in clinic. Methods 3-dimensional finite element analysis models of Tapered,Square and Ovoid arch form maxillary based on the MBTTM Arch Form Templates were constructed by using Solidworks software.The cortical bone thickness was defined as 2 mm at all points.The implants with 13 mm in length were inserted between the mental foramina.The 2 posterior implants were set at 4 inclination angle,from 0°to 45°.All implants were splinted with superstructure.A 100 N pressure was applied to 2 kinds of loading positions on the superstructure models.The maximum Von-Mises stress on cortical bone around the implants was measured by using Abaqus software.Results The maximum Von-Mises value in maxillary cortical bone with All-on-4 implants in 3 arch forms showed significant differences.In Square model the concentration of stress was advanced with the increasing of implant inclination angle;In Ovoid model,the maximum Von-Mises value was observed at 15°in posterior region,and was increased significantly by pasted 15° in anterior region;In Tapered model,the Maximum Von-Mises value was increased greatly at 45°.Conclusion Arch forms have appreciable impact on stress distribution of peri-implant.In Square arch form,the inclination of implant should be avoided;in Ovoid arch form,the inclination angle should be under 15°;in Tapered arch form,the inclination should be under 30°.

Abstract:Objective To study the influence of tripterygium glycosides (LGTDG) in the bone morphogenetic protein(BMP) signal transduction pathway and BMP-2 expression,and to clarify the mechanism of anti-ankylosing spondylitis (AS) ossification of LGTDG.Methods The in vitro 　cultured AS fibroblasts were divided into control group and 0.5,1.0,1.5,2.0 mg/L LGTDG groups.The alkaline  phosphatase (ALP) activities of the cells and optimal drug concentrations in various groups were detected;CCK-8 assay was used to detect the proliferation rate of the cells in 1.0 mg/L LGTDG group;the biochemical tests were performed to quantitatively detect the BMP-2 expression levels in control group and 1.0 mg/L LGTDG group;Western blotting method was used to determine the    BMP-2 and Cbfal protein expressions.Results The ALP activities in LGTDG groups were lower than that in control group, especially in 1.0  mg/L LGTDG group (P<0.01).The CCK-8 assay results showed that the proliferation rate of  AS fibroblasts in 1.0 mg?L-1LGTDG group was significantly low than  that in control group(P<0.01),the proliferation rate reached the peak at the 4th day  after LGTDG treatment and entered into the plateau phase,then the proliferation rate of the cells was decreased.The biochemical assay and Western blotting results indicated that the protein expression levels of BMP-2 in 1.0 mg/L LGTDG group was significantly lower than that in control group (P<0.01).Conclusion LGTDG can effectively inhibit the BMP-2 expression in AS fibroblasts and delay the cells to differentiate into the osteoblasts and lead to AS ossification by BMP signal transduction pathway.

Abstract:Objective To investigate the effect of somatostatin on the electrophysiological changes of early diabetic rats,and to clarify its therapeutic effects on retinopathy in early diabetic rats. Methods 20 Wistar male rats,which were induced into diabetic rat models by intraperitoneal injection of streptozotocin (STZ),were randomly divided into somatostatin therapy group and model control group,10 in each group. The rats in somatostatin therapy group were injected with somatostatin 10 μg/kg/d,for 8 weeks;the rats in model control group received the same volume of saline. Another 10 male Wistar rats were used as normal control group. The changes of blood glucose and electroretinogram were measured,and the indicators were used for statistical analysis. Results Compared with model control group,the blood glucose was declined in somatostatin therapy group at the 8th week (P<0.05). Compared with normal control group,the latent periods of a-wave,b-wave of rod-response and max-response were significantly extended in somatostatin therapy group and model control group at the 1st,2nd,4th,6th,and 8th week (P<0.05). The latent periods of a-wave,b-wave of rod-response and max-response in somatostatin therapy group were shorter than those in model control group at the 8th week (P<0.05). Compared with model control group,the amplitude had no changes at each observation time point in somatostatin therapy group (P>0.05). Conclusion Somatostatin has a therapeutic effect on early diabetic retinopathy in rats.

Abstract:Objective  To explore the effects of trimetazidine on myocardial free radical injury induced by pirarubicin， and to clarify the protective effect and mechanism of trimetazidine on myocardial injury induced by pirarubicin.Methods 36 Wistar rats were randomly divided into pirarubicin group（n=13），trimetazidine intervention group（n=13） and control group （n=10）.The rats in pirarubicin group and trimetazidine intervention  group were injected with pirarubicin 2.5 mg/kg by the vena caudal once a week for 6 weeks.The rats in trimetazidine intervention  group were intragastricly infused with trimetazidine 5.4 mg/kg/d one day for 8 weeks before making the model. At the end of the experiment,the malonaldehyde (MDA) level，nitrogen oxide（NO） level，superoxide dismutase(SOD) activity，and nonprotein sulfhydryl (NPSH) level in  myocardium tissue were measured. The histological changes of myocardium tissue were detected by electron microscope. Results Compared with control group ,the levels of MDA and NO in pirarubicin group were increased（P<0.05），and the SOD activity and NPSH level in pirarubicin  group were decreased（P<0.05）.Compared with pirarubicin  group,the levels of MDA and  NO in trimetazidine intervention   group were decreased（P<0.05）， the SOD activity  and NPSH level in trimetazidine intervention  group were increased（P<0.05）.Under  electron microscope,the myocardiocytes of the rats in pirarubicin group showed irregular arrangement in sacromere structure,shrinkage in nuclear membrane,vacuolation in nuclear matrix,obvious mitochondria swelling,deposition of metachromatin throughout the nucleus,and an indistinct view of intercalated disc with isolation;while in trimetazidine intervention group the nucleus was round and nuclear membrane was indented,myofilament bundles were decreased slightly with a regular arrangement，intercalated disc oriented transversely with partial vague in cell junction structure,and mitochondria slightly swelled.Conclusion Trimetazidine has the protective effects on the damaged myocardiocytes caused by pirarubicin,and its mechanism may be related to reducing the production of free radicals and decreasing the injury of structures within the cells,such as the nucleus,mitochondria and intercalated disc.

Abstract:Objective To investigate the mechanism of increasing of the level of kidney injury molecule-1(KIM-1) in culture supernatant of human kidney cells(HKC) induced by cyclosporine A(CsA),and to clarify the relationships between the expression levels of KIM-1 and p38 MAPK pathway and  ERK1/2MAPK pathway in HKC. Methods The HKC at logarithmic growth phase were randomly divided into control group,CsA control group,CsA + p38 kinase inhibitor group,p38 kinase inhibitor group,CsA + ERK1/2 inhibitor group and ERK1/2 kinase inhibitor group. The inhibitory rates of proliferation of HKC in various groups were  detected by MTT assay,and the expression levels of KIM-1 in HKC supernatant in various groups were detected by ELISA;the survival rates,apopototic rates and necrotic rates  of the HKC in various groups  were detected by flow cytometry. Results Compared with control group,the expression level of KIM-1 protein in the supernatant of HKC in CsA control group was significantly increased (P<0.05),and the survival rate was significantly decreased (P<0.05),while the apoptotic rate and the necrotic rate were significantly increased (P<0.05).Compared with control group,the survival rates，the apoptotic rates and the necrosis rates of cells in p38 kinase inhibitor group and ERK1/2 kinase inhibitor group had no significant differences(P>0.05). Compared with CsA control group,the expression levels of KIM-1 protein in CsA + p38 kinase inhibitor group and CsA + ERK1/2 kinase inhibitor group were significantly decreased (P<0.05),and the survival rate was significantly increased (P<0.05),while the apoptotic rate and the necrotic rate were significantly decreased (P<0.05). Conclusion p38 MAPK pathway and ERK1/2MAPK pathway are involved in the process of up-regulation of the KIM-1 level in HKC culture supernatant induced by CsA,and the expression of KIM-1 may become the biochemical marker of clinical monitoring of CsA nephrotoxicity.

Abstract:Objective To investigate the effects of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) on the gingival wound healing，and to lay a foundation for its application in the field of oral cavity.Methods 16 healthy rabbits were randomly divided into experiment and control groups(n=8).2　mm upper and lower incisors’ labial gingivae were removed by scalpel.After operation,rhGM-CSF gel was applied to the surface of the wound gum in experiment group,while saline instead of drugs to the control group.2 rabbits were executed at 3,7,11,and 15 d after operation.HE staining was performed to observe the epithelium and epithelial connective tissue structure,distribution,and the changes of various cells of the rats;the  number of positive epithelial cells and fibroblasts were observed by PCNA dyeing.Results On the 3rd day, the  inflammatory cell infiltration was found in two groups,but a greater number of the cells were observed in experiment group; neovascularization was seen in both groups on the 7th day,and the fibroblasts and collagen fibers were observed too,but the extent of neovascularization in experiment group was more significant.On the 11th day,the fibroblasts proliferation was seen in two groups,but the extent in experiment group was more widely; on the 15th day,the trauma of gingivae of the rabbits in two  groups returned to normal,there was no significant difference between two groups.Over time,the number of epithelial cells in proliferation was gradually increased,reached the peak on the 15th day.The number of fibroblasts began to increase from the 3rd day and reached the peak on the 11th day,and significantly reduced to the lowest value  on the 15th day.The number of proliferative epithelial cells in experiment group was significantly higher (P<0.01)on the 3rd day and the 11th day than those in control group(P<0.01);the number of proliferative  fibroblasts was significantly higher (P<0.05) on the 11th day and the 15th day than those in control group(P<0.05),but there were no significantly statistical differences between two groups at other time pionts(P>0.05).Conclusion  RhGM-CSF can promote the infiltration of inflammatory cells,angiogenesis, proliferation of epithelial cells and fibroblasts in gingivae.RhGM-CSF is beneficial with wound healing on gum tissue.

Abstract:Objective  To copy the mouse aging model with D-galactose,and to investigate the role of Schisandra total lignin (SCL) in the  mouse brain tissue aging and its mechanism.Methods 50 mice were radomly divided into control group,model group (100 mg?kg-1?d-1),low dose (35 mg/kg/d) of SCL group (SCL-L)， middle dose  (70 mg/kg/d) of SCL group (SCL-M) and high dose (140 mg?kg-1?d-1) of SCL group (SCL-H)（n=10）.D- galactose ( 100 mg?kg-1?d-1 ) was injected into the mice hypodermically for 10 weeks to induce aging models in all the groups except control group,and 35,70, and 140 mg/kg/d  SCL were administered for 10 weeks in SCL groups.The learning and memory abilities were measured by the Water Maze test.The expression levels of Bax,Bcl-2,ubiquitin (Ub),microtubule-associated protein light chain 3 (LC3) in the brain tissue of  the mice in various groups were observed by Western blotting method.The LC3 protein expressions in mouse brain cortex and hippocampus were observed by immunohistochemistry.Results  In learning and memory test,compared with control group,the swimming time of the mice in model group was increased (P<0.05),and the number of errors was increased (P<0.05);compared with model group,the swimming time in SCL-L,SCL-M and SCL-H groups was decreased (P<0.05) and the number of errors was also decreased (P<0.05). Compared with control group，the expression level of Bax was increased (P<0.05),the expression level of Bcl-2 was decreased (P<0.05),the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were increased (P<0.05) in model group;compared with model group,the expression level of Bax was decreased (P<0.05),the expression level of Bcl-2 was incerased (P<0.05),and the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were decreased (P<0.05) in SCL-L,SCL-M and SCL-H groups.In control group,the neuronal morphology was normal,and none of brown granules were visible in the cytoplasm of mouse brain cortex and hippocampus and the expression of LC3 protein was negative.In model group,the neurons were degeneration,and the number of LC3 protein positive cells in the cerebral cortex and hippocamptal tissue was increased (P<0.05).In SCL-L,SCL-M and SCL-H groups,the number of degenerative neurons was decreased,and the number of LC3 protein positive cells was decreased (P<0.05).Conclusion SCL can inhibit the D-galactose-induced brain tissue aging in the mice,and the mechanism is related to regulating autophagy and inhibiting apoptosis.

Abstract:Objective To investigate the damage of hippocampal neurons induced by  chronic cerebral ischemia in the rats,and to clarify its mechanism.Methods 90 healthy adult male SD rats were randomly divided into sham operation group (n＝30) and experimental group (n＝60).The chronic cerebral ischemia rat models were established by permanently ligating the common carotid arteries on both sides of the rats in experimental group.The rats in sham operation group were established by incising the  cervical median,only the common carotid arteries on both sides were separated without ligating.The rats in sham operation group and experimental group were respectively sacrificed at the  7th,14th and 21st  day after operation.At each time point 10 rats in sham operation group and 20 rats in experimental group were selected and sacrificed.Immunohistochemistry staining was used to observe the dynamic changes of the expression levels of choline acetyltransferase(ChAT) in hippocampus neurons,and TA-FE method was used to observe the dynamic changes of hippocampal microvascualr architecture.MiVnt image analytical system was used to quantitatively analyze the immunohistochemistry result,the microvessel density (MVD) and micorvessel area density (MVA) of horizontal part of hippocampus in the rats.Results Compared with sham operation group,the ChAT expression levels in hippocampus neurons of the rats in experimental group at different time points were significantly decreased（P<0.05);and with the prolongation of time,the ChAT expression levels were gradually decreased;the ChAT expression level in 14-day experimental group was significantly lower than that in 7-day experimental group（P<0.05);the ChAT expression level in 21-day experimental group was significantly lower than those in 7-day and 14-day experimental groups（P<0.05).The MVD and MVA of hippocampus of the rats in experimental group at different time points were obviously decreased compared with sham operation group（P<0.05）;the MVA was gradually decreased with the prolongation of time,and the MVA of hippocampus of the rats in 21-day and 14-day experimental groups were obviously decreased compared with 7-day experimental group（P<0.05);the MVD was gradually decreased with the prolongation of time,the MVD of hippocampus of the rats in 21-day and 14-day experimental groups was obviously decreased compared with 7-day experimental group（P<0.05);the MVD of hippocampus of the rats in 21-day experimental group was obviously decreased compared with 14-day experimental group（P<0.05).Conclusion Chronic cerebral ischemia can lead to the progressive decrease of the ChAT expression level,MVD and MVA of hippocampus of the rats  to aggravate gradually the learning and memory dysfunction,which may be one of the reasons of vascular dementia.

Abstract:Objective To explore the expression of transient receptor potential channel 6(TRPC6) in human breast cancer cells, and to clarify the correlation of TRPC6 with  the invasion potential of breast cancer cells.Methods The human breast cancer cell strains MCF-7(hypo-invasion group) and MDA-MB-231(hyper-invasion group)  were cultured.The expressions of TRPC6mRNA and protein in in two groups were detected by RT-PCR and Western blotting methods.Then the MDA-MB-231 cells were divided into control group and SKF96365 group，the effects of SKF96365 on the invasion ability of MDA-MB-231 cells in vitro were explored by wound healing assay and Transwell experiment.Results The results of Western blotting and RT-PCR showed that the  expression  levels of TRPC6 mRNA and  protein   in MDA-MB-231 cells were higher than that in MCF-7 cells（P<0.05).The wound healing assay showed the numbers of migrating cells in 5,25 and 40  μmol/L SKF96365 groups (76.24±7.54，45.33±4.50，25.12±1.57) were lower than those in control group (130.48±9.55) （P<0.05).The Transwell experiment results indicated that the invasiveness of MDA-MB-231 cells were inhibited significantly by SKF96365 compared with control group (P<0.05).Conclusion  The invasion ability of human breast cancer MDA-MB-231 cells is promoted by upregulating the TRPC6 expression，which indicates that the TRPC6 may play role in the metastasis of human breast cancer.

Abstract:Objective To investigate the expression   of anoctamin 1 in Chinesehamster ovary cells (CHO)and to analyze the functional properties of its ion channel, and to provide experimental basis for study on the physiological function of calcium-activated chloride channel. Methods The whole sequence of anoctamin 1 was obtained by PCR technique and was subcloned into pcDNA3.1 to construct the expression vector pcDNA3.1-anoctamin 1  was transfected into CHO by liposome-mediated transfection and the CHO stably expressing anoctamin 1 were aquired by selection with zeocin. The expression and distribution ofanoctamin 1 in CHO were measured by RT-PCR technique and inverted fluorescence microscope.The functional properties of anoctamin 1 were measured with halide-sensitive fluorescence proteins YFP-H148Q/I152L. The PBS buffer solution with calcimycin and high concentration of iodine ion wasused as experimental group,andthe PBS buffer solution without  calcimycin and high concentration of iodine ion was used as  control group.Results  The results of digestion and sequencing confirmed that anoctamin 1 was cloned into pcDNA3.1 by electrophoresis and blast.The results of RT-PCR and  inverted fluorescence microscope indicated that anoctamin 1 was expressed in CHO.The results of I-  influx as measured by halide-sensitive fluorescence proteins YFP-H148Q/I152L showed that anoctamin 1 had the more functionalproperties of trans-epithelial transporting I－,and the transporting speed in experimental group was higher than that in control group(P<0.05). Conclusion Anoctamin 1 can be expressed highly in the CHO；Anoctamin 1  expressed in CHO has the characteristics of calcium-activated chloride channel.

Abstract:Objective To measure the vitamin B1 levels in plasma,erythrocytes an
d urine of the patients with type 2 diabetes,and to analyze the correlation of  vitamin B1 level with the progression of diabetic nephropathy,and to clarify the metabolism of nutrtion mechanism of type 2 diabetic nephropathy.Methods Total 90 patients with type 2 diabetes and 30 healthy volunteers were recruited.According to the levels of microalbuminuria,the patients with type 2 diabetes were divided into non diabetic nephropathy (NDN) group,early diabetic nephropathy (EDN) group and clinical diabetic nephropathy (CDN) group (n=30);the healthy people was used as normal control (NC) group.The vitamin B1 levels in the plasma,erythrocytes and urine were examined with high performance liquid chromatography (HPLC).The correlation of microalbuminuria with the  plasma viamin B1 level was analyzed.Results The level of vitamin B1 in plasma of the patients in NC group was (7.1±3.3)  μg/L,while it was (4.0±2.3)  μg/L in NDN group,(3.1±1.0)  μg/L in EDN group and (2.3±0.6)  μg/L in CDN group.Compared with NC group，the vitamin B1 levels in the plasma in NDN,EDN and CDN groups dropped 43.7%,56.3%， and 67.6%, respectively (P<0.05). The excretion of vitamin B1 in urine were (2.9±0.8),(9.0±4.7),（11.7±3.9）,and　（15.6±5.0） μg/L in NC group,NDN group,EDN group and CDN group, respectively.Compared with NC group，the vitamin B1 levels in the urine in NDN,EDN and CDN groups were increased by 2,3 and 4 times, respectively (P<0.05).A negative correlation was found between the level of microalbuminuria and the level of vitamin B1 in plasma (r=－0.62,P=0.013).Conclusion  Vitamin B1 deficiency can be observed in the early stage of diabetic nephropathy,and the level of vitamin B1 is closely correlated with the progression of diabetic nephropathy.

Abstract:Objective To investigate the expressions of miR-21 in carcinoma tissue and serum of the patients with papillary thyroid carcinoma (PTC),and to clarify its clinical significance.Methods Real-time PCR was used to detect the expression levels of miR-21 in carcinoma tissue and serum of 46 PTC patients,thyroid tissue and serum of  35 cases of nodular goiter and 18 healthy people.The correlations of miR-21 expression level with the clinicopathological features of PTC patients were analyzed.Results The relative expression levels of miR-21 in carcinoma tissue and serum of the PTC patients were 2.74±1.53 and 0.21±0.08,and they were 0.92±0.81 and 0.08±0.05 in carcinoma tissue and serum of the patients with nodular goiter,0.86±0.39 and 0.07±0.03 in normal papillary thyroid tissue and serum of 18 healthy people.The expression levels of miR-21 in carcinoma tissue and serum of the patients with PTC were higher than those of the patients with nodular goiter and healthy people (P<0.05).The expression level of miR-21 in serum and carcinoma tissue of PTC patients had a positive correlation with the extraglandular invasion,distant metastasis,lymph node metastasis, and TNM stages (P<0.05);and it had no correlation with gender and tumor sizes.Conclusion The expression levels of miR-21 in carcinoma tissue and serum of the PTC patients are increased.The up-regulated expression of serum miR-21 is correlated with the clinicopathological features of PTC patients,which indicates that miR-21 could be used as a potential marker for early detection of PTC.

Abstract:Objective To understand the characteristics of surface electromyography（sEMG）of vastus medialis muscle fatigue induced by isokinetic flexion and extension movement,and to further provide references for the application of sEMG on dynamical movements.Methods Arial isokinetic equipment and ME6000 equipment were combined to analyze the time domain indices including root mean square (RMS) integrated electromyography (iEMG) and spectrum indices including median frequency (MF) and mean power frequency (MPF) for vastus medialis from ten students from Shenyang Sport University during isokinetic fatigue.Results During the isokinetic muscle fatigue the RMS and iEMG were significantly increased (P<0.05),accompanying with higher effect sizes for  iEMG than for  RMS. As for the spectrum indices,MF and MPF were significantly decreased (P<0.05) with larger effect sizes for MPF than for MF.The increase rate of iEMG was significantly higher in iEMG than RMS (P<0.05), and the decline rate of MPF was larger than MF (P<0.05).Conclusion The iEMG as a time domain index and MPF as a spectrum index can be sensitive indexes to assess the dynamic muscle fatigue.

Abstract:Objective  To study the features and differences of electromyography of diabeticpolyneuropathy (DPN) and alcoholic peripheral neuropathy (APN),and to provide reference basis for the clinical application of electromyography.Methods 58 patients with DPN and 30 patients with APN were used as subjects.Nerve conduction studies (NCS) and sympathetic skin response (SSR) were performed in the patients,all data were analyzed.Results In the patients with DPN,the abnormalities  of NCS and SSR were increased with the prolongation of the time diabetes,and the abnormality of  SSR was higher than that of NCS(P<0.05).In the patients with APN,both demyelination and axonal loss in motor and sensory nerves were significantly involved,and the abnomalities of NCS and SSR were higher than those of the DPN patients (P<0.05 or P<0.01);but the proximal nerves were just involoved mildly.Conclusion Both DPN and APN have characteristic electrophysiological features.Early electromyography is useful for the early diagnosis of DPN and APN.According to the electrophysiological features of DPN and APN, the reason of peripheral neuropathy in the patients who have diabetes mellitus and alcoholism could be differentiated.

Abstract:Objective To detect the serum levels of carbohydrate antigen 199(CA199),carbohydrate antigen 125(CA125) and carcino-embryonic antigen(CEA) in different population， and to explore their significances in diagnosis of pancreaticcancer.Methods The serum levels of CA199,CA125 and CEA in 56 patients with pancreatic cancer(pancreatic cancer group),56 patients with benign pancreatic disease(benign pancreatic disease group) and 56 healthy volunteers（normalcontrol group）were detected,then the positive rates of CA199,CA125 and CEA and the sensitivities and specificities of CA199,CEA and CA125 for the diagnosis of pancreatic cancer were compared among three groups.Results The levels of serum CA199,CA125 and CEA of the patients in pancreatic cancer group were significantly higher than those in benign pancreatic disease group and normal control group (P<0.05).The positive rates of CA199,CEA and CA125  in pancreatic cancer group were 87.50 %,71.42%，  and 60.71%,respectively. The  positive rates of the three tumor markers of the patients in pancreatic cancer group were obviously higher than those in other two groups (P<0.05).The sensitivity of CA199（87.50%） was the highest,while the specificity of CEA （91.96%） was the higest.The method of parallel combined testing increased the diagnostic sensitivity to 95.12% and negative predictive value to 90.47%,but its specificity and the positive predictive value were decreased.Serial combined examination increased the diagnostic specificity to 92.86% and the positive predictive value to 89.19%,but its sensitivity and negative predictive value were decreased.Conclusion Combined detection of serum levels of CA199,CA125 and CEA has higher sensitivity and specificity than single-marker detection in the diagnosis of pancreatic cancer.

Objective  To study the expressions of casein kinase 2α(Ck2α),β-catenin,survivin in ovarian cancer tissue and analyze the relationships between them, and to explore the clinical application value.Methods Immunohistochemistry staining  was used to detect the expressions of Ck2α,β-catenin and survivin in 8 cases of simple ovarian cysts tissue,8 cases of ovarian benign tumor tissue and 29 cases of ovarian cancer tissue;  the correlations   of  the expressions of Ck2α,β-catenin,survivin in ovarian cancer tissue andtheir  associations with the clinicopathologic characteristics were analyzed.Results The expression
levels of Ck2α, β-catenin and survivin in ovarian cancer tissue were significantly higher than those in simple ovarian cysts  tissue and ovarian benign tumor tissue(P<0.05);the Ck2α expression level   in ovarian cancer tissue  was positively corelated with  the expression levels of β-catenin and survivin
(r=0.438,r=0.479,P<0.05);the expression levels of Ck2α,β-catenin and survivin in  low-medium differentiation group were significantly higher
 than those in high differentiation group (P<0.05);the expression levels of Ck2α,β-catenin and survivin in stagesⅡand Ⅲ group
 were significantly higher than those in  stage Ⅰ group(P<0.05).Conclusion The protein expression levels of Ck2α,β-catenin and survivin are  increased in ovarian cancer tissue,and three are positive corelations between them;Ck2α,β-catenin and survivin  may play an important role in the occurrence and development of ovarian cancer;the combined detection of them has important clinical value.
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Objective To investigate the expressions of inflammatory factors in papillary thyroid carcinoma tissue and their relationships with the clinical pat
hological characteristics,and to provide an experimental basis for guiding clinical treatment and prognosis.Methods 74 patients with papillary thyroid carcinoma dignosed by puncture were selected as experimental group,and  26 cases of healthy people were used as normal control group.The fasting venous blood samples  from the subjects in two groups were collected,and the postoperative specimens from cancer tissue,adjacent normal tissue and the fasting venous blood 7 d after operation of the patients in experimental group were collected again.The interleukin 1α (IL-1α),interleukin 1β (IL-1β) and cyclooxygenase -2 (COX-2) protein expression levels in serum,cancer adjacent normal tissue and cancer tissue were detected by ELISA;the gene and protein expression levels of IL-1α,IL-1β and COX-2 were examined by real-time fluorescent quantitative method and immunohistochemistry;th
e relationships between their expressions and clinical stages,pathological types, lymph node metastasis of thyroid papillary carcinoma were analyzed.
Results The expression levels of serum IL-1α,IL-1β and COX-2 protein of the patients in experimental group  were significantly higher than those in normal
 control group(P<0.01);the gene and protein expression levels of IL-1α,IL-1β and COX-2 in thyroid papillary carcinoma tissue were significantly
 higher than those in adjacent tissue (P<0.01).There were  positive correlations between the expressions of IL-1α and IL-1β,IL-1α and COX-2,IL-1β and
 COX-2 in thyroid papillary carcinoma tissue (r=0.64,P=0.035;r=0.71,P=0.042;r=0.69,P=0.038).Conclusion The expression levels of I
L-1α,IL-1β and COX-2 in thyroid papillary carcinoma tissue are increased,suggesting that inflammation is involved in metastasis and invasion  of thyroid papillary carcinoma,its application is expected to become a new means of cancer treatment strategies.

Objective To investigate the features of empathy deficiency of the patients with schizophrenia,and to analyze the relationship between empathy ability,clinical symptoms,and social function.Methods The patients with first-episode  schizophrenia (n=30) and the normal controls (n=30) were selected. Interpersonal Reactivity Index-C( IRI-C),Positive and Negative Syndrome Scale (PANSS),and the Scale of Social Function in Psychosis Inpatients (SSPI ) were used to measure the score of empathy,mental symptoms and the social ability;the total score of empathy and the scores of four scales in two groups were compared;the relationships between the empathy ability score and clinical symptom score,social function score in patients group were analyzed.
Results The IRI-C score of the patients in schizophrenia patients group was lower than that in normal control group (P<0.05).The schizophrenia patients with higher PANSS negative syndromes had the lower IRI-C score,and serious empathy deficiency;there was negative correlation between empathy ability and PANSS negative syndrome in schizophrenia patients group（r=－0.572，P<0.01）;the IRI-C score wasn’t  correlated with SSPI score（r=0.166，P>0.05） in schizophrenia patients group.Conclusion The first-episode schizophrenia patients have empathy deficiency,which is affected by the negative syndromes.The more severe negative symptoms  of the  patients are， the worse empathy ability is.

Objective To investigate the surgery method for early spontaneous posterior interosseous nerve entrapment,to observe the postoperative efficacy in tr
eatment,to evaluate the surgical outcomes,and to provide a foundation for clinical choice of reasonable operation scheme.Methods 21 cases of early spontaneous posterior interosseous nerve entrapment received operation.13 cases (18 arms) were treated by neurolysis,and 8 cases (8 arms )　were treated by nerve grafts with small vessels wrapping operation;all the patients were followed up for 10 to 20 months;the finger extensor muscle strength and metacarpophalangeal joint activity were evaluated.Results The lesion extensor muscles were part of the denervation changes.Neurolysis muscle strength:14 arms were excellent(82.35%),2 arms were good,1 arm was fair,1 case was lost;extensor function:15 arms were excellent(88.24%),1 arm was good,1 arm  was fair,1 case was lost.Nerve graft  muscle strength:6 arms were excellent(75.00%),1 arm was good,1 arm was fair;extensor function:7 arms were excellent(87.50%),0 arm was good,1 arm was fair. Conclusion Neurolysis can have a good efficacy in treatment of early spontaneous posterior interosseous nerve entrapment without obvious degeneration.When the severe degeneration of entrapment nerve happens,a nerve graft surgical treatment is needed.

Objective To investigate the difference  of curative effects between the complete video assisted thoracic surgery (C-VATS) and traditional open surgery
 in the treatment of elderly patients with lung cancer, and to clarify the specially curative effect of C-VATS.Methods 80 patients with lung cancer without diabetes mellitus,chronic brouchitis and heart diseases aged over 65 years were enrolled in the study.They were divided into  C-VATS group and open surgery for lobectomy (OPEN) group (n=40) according their wishes. The operation time,number of lymph nodes removed,amount of intraoperative blood loss,postoperative thoracic drainage,chest tube duration,postoperative hospitalized time,postoperative pain degree,incidence of postoperative complications,preoperative and postoperative sedentary heart rate (HR) of the patients in two groups were compared.Results The operation time in C-VATS group(193.12 min±59.06 min) was longer than that in OPEN group(167.17 min±54.01 min)(P<0.05),and the number of lymph nodes removed in C-VATS group(14.6±7.5) had no significant difference compared with OPEN group(15.2±4.5)(P>0.05).The postoperative pain degree in C-VATS
 group(2.54±0.12) was lower than that in OPEN group(4.61±0.10)(P<0.05);the postoperative chest rube duration in C-VAS group(6.14 d±3.32 d) was shorter than that in   OPEN group  (11.67 d±4.13 d);the postperative hospitalized time in C-AVA group(8.52 d±3.25 d)was shorter than that in OPEN group

(14.76 d±6.11 d)(P<0.05).There were no significant differences of intraoperative blood loss(231.28 mL±203.15 mL vs 213.33 mL±187.18 mL),postoperative thoracic draginage(1 597.83 mL±1 049.29 mL vs  1 690.68 mL±1 043.37 mL),incidence of postoperative complications between C-VATS group and OPEN group(P>0.05).The postoperative sedentary HR in C-VATS group and  OPEN group were significantly higher than preoperative;the  postoperative sedentary HR recovered  to the preoperative level 3 d after operation in C-VATS group and it  recovered to the preoperative level  7 d after operation  in OPEN group.Conclusion Compared with traditional open thoracic surgery, C-VATS    owns the characteristics such as less trauma,less pain
,less hospital stay and better postoperative recovery in the aged patients with lung cancer.
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Objective To compare the clinical effects of laparoscopic Nissen fundoplication(LNF)combined with highly selective vagotomy(HSV) (LNFHSV)in   treatment of gastroesophageal reflux disease(GERD),and to provide reference for the clinical application of LNFHSV.Methods 22 patients with a history of GERD unrelieved by medication and underwent LNFHSV were selected.At the same time ,36 patients with GERD underwent LNF were used as control.The  mean operation time,hospital stay,incidence of main operative complications,HSS complete remission and  Demeester scores of the patients in two groups were retrospectively analyzed.Results The Demeester scores,hospital stay,incidence of main operative complications of the patients in two groups before operation had no significant differences（P>0.05）.The mean operation time in LNFHSV group (90 min±35 min）was longer than that in LNF group（65 min±21 min）.The Demeester scores of the patients in two groups after operation had significant difference（P<0.05）.
The HSS complete remission rate of the patients in LNFHSV group was 91.1%,the part remission rate was 8.9%,and no effectiveness was  0;they were 83.3%，13.8%，and 2.8% in LNF group;there were significant differences between two groups（P<0.05）.Conclusion LNFHSV has better effectiveness in controlling GERD than LNF procedure.

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Objective To investigate the application of vascularized free fibula flaps in reconstruction of mandibular defect,and to evaluate the survival rate and re
pair effect of the method.Methods 16 patients with mandibular tumor,having a desire to reconstruct mandible,and their systemic state could tolerate the long time operation,were selected to reconstruct mandibular defect by vascularized fibula flaps，of which 12 were male,4 female,aged 24-66 years old,average 45.2 years old.The resection of primary tumor and free fibula flaps were simultaneously conducted,then the  fibular flaps were shaped according to the defect location and size of the mandible,and were fixed with reconstructive titanium plate for repair and reconstruction of mandible.The survival of flaps was determined by skin color,texture,and skin temperature of the flaps.The reconstruction effects were evaluated through the patients’ surface like photos and X-ray picture of mandible before and after operation.Results All of flaps were survived and no serious complications were found.The complications of the supplied sites were skin tension and wound dehiscence,which were healed by dressing.The mandibular reconstruction effects were good through 2 or more persons’ double blind evaluation.Conclusion Free fibula flaps have high survival rate and good results in   mandibular reconstruction,and they can meet the needs of various types of mandibular defect repair.

Objective To discuss the efficacy and safety of triamcinolone acetonide (TA) retrobulbar injection combined with panretinal photocoagulation (PRP) i
n the treatment of diabetic macular edema (DME),and to explain the advantages of this combination treatment method in treating DME.
Methods Sixty-four cases (64 eyes) with macular edema due to diabetic retinopathy(DR)  were randomly divided into combination treatment group(n=34,treated with TA retrobulbar injection combined with PRP) and PRP group(n=30,received PRP treatment only).The changes
 in the best corrected visual acuity (BCVA),the fovea centralis thickness (CMT),and the intraocular pressure(IOP) before treatment and in 1 month,3 months,6 months after treatment were compared.  Results The BCVA was improved in both groups after treatment.However,the BCVA improvement in combination treatment group was statistically more significant (P<0.05) compared with PRP group in 1 month,3 months and 6 months after treatment.The resolution of DME after treatment as compared with that before treatment,the CMT reduced in two groups in 1 month,3 months,6 months after treatment.At each stage,the CMT in  combination treatment  group was lower than PRP group (P<0.05).No complications of glaucoma，cataract,vitreous hemorrhage,retinal detachment,choroidal detachment,and endophthalmitis were  found in all patients.Conclusion For the patients with DME,the combination
 treatment with TA retrobulbar injection and PRP has a better therapeutic effect than the sole PRP treatment in reducing the macular edema and improving the BCVA.The operation is simple and has less complication.
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Objective To compare the diagnostic results of 18F-deoxyglucose positron emission tomographic-computed tomographic (18F-FDG PET-CT) imaging ,CT and golden standards in TNM stating of non-small cell lung cancer (NSCLC),and  to  evaluate the diagnostic value of 18F-FDG PET-CT imaging in TNM staging of NSCLC.Methods 18F-FDG PET-CT was performed in the 65 patients with NSCLC who were diagnosed by pathology.Blind interpretation was used to dignose the TNM staging.The sensitivity(Se),specificity(Sp),positive predict value(PPV) and negative predict value(NPV),coincidence rate and Kappa analysis were performed to evaluate the values of PET-CT and CT in diagnosing TNM staging of NSCLC.Results Compared with golden standard,the di
agnostic results of T staging among 65 NSCLC patients by CT and PET-CT were respectively as coincidence rate=72.31%,Kappa=0.46;coincidence rate=78.46%,Kappa=0.60;the diagnostic results of N staging among 58 NSCLC patients by CT and PET-CT were respectively as coincidence rate=74.14%,Kappa=0.50;coincidence rate = 87.94%,Kappa=0.78;the diagnostic results of lymph node metastasis by CT and PET-CT were respectively as Se=64.04%，Sp=77.53%，PPV=85.07%，NPV=51.88%，coincidence rate =68.54%，Kappa=0.37；Se=91.01%，Sp=83.15%，PPV=91.53%，NPV=82.22%，coincidence rate =88.39%，Kappa=0.74;the total detection rate and amount of metastasis by PET-CT were significantly higher than that by CT.Conclusion ¹⁸F-FDGPET-CT  has significant value on the diagnosis of  the TNM staging of NSCLC.

Objective To determine  the biomechanical parameters of left ventricular by using  velocity vector imaging(VVI),and  to indirectly assess the  coron
ary artery stenosis  with  VVI in the patients with  coronary artery disease(CAD).Methods 52 patients  who had one coronary artery lesions at least diagnosed
by coronary angiography (CAG)  were divided into coronary artery mild stenosis group and severe stenosis group;21 patients in mild stenosis group had one coronary stenosis <75%;31 patients in  severe stenosis group had one coronary artery stenosis ≥ 75% at least.At the same time,20 cases of normal people without coronary artery stenosis showed by CAG were selected as normal control group.VVI  was used to  detect  the  left ventricular wal
l segments of the overall longitudinal strain (GLS),the overall circumferential strain (GCS) and the overall radial strain (GRS).Results
The absolute values of GLS,GRS,GCS of the patients in  mild and severe coronary artery stenosis groups were significantly decreased  than those in  normal control group(P<0.05),and the strain parameters in severe stenosis group were  decreased more significantly,there  were
significant differences compared  mild stenosis group  (P<0.05).The  distribution of the segments with decreased longitudinal strain  matched  t
he LV myocardial segment with the coronary stenosis rate ≥ 75%,and GLS had  the most sensitivity.TheGLS   in normal control group,mild stenosis group and severe stenosis group were negatively correlated to left ventricular ejection fraction (LVEF) (r=－0.58,P<0.05;r=－0.51,P<0.05;r=－0.43,
P<0.05).GLS-16.14 %  was used to  assess  the  severe coronary artery stenosis with requiring  the implementation of PTCA treatment  as the diagnostic cut-off point with  sensitivity 96.8%,specificity of 70%,the highest Yuedden index  0.668. Conclusion The decreasing of left ventricular strain could be detected by VVI,which  suggests that   severe coronary artery stenosis exists in coronary artery;the distribution of the segments with significantly decreased strains
    can be used to assess the  coronary lesions and stenosis degrees.

Objective
To study the awareness of Brucellosis prevention knowledge and prevalence of risk behaviors among the primary and secondary school  students from  breeding livestock’s families in Western pastoral areas of Jilin province,and to provide evidence for the prevention and control of Brucellosis.Methods With application of multi-stage random sampling method,a questionnaire survey which including Brucellosis prevention knowledge and  risk behaviors was conducted among 209 primary and  secondary school students in west part of Jilin province in 2012.Results 105（50.97%） students had heard Brucellosis among 206 students,the older students knew more about Brucellosis than the younger（χ2=28.80，P=0.00）.The M(Q) of the score of the Brucellosis-related knowledge was 8（6.00,9.00）,the total passing ratewas 28.64%.72(68.61%) students learned Brucellosis from village doctors and acquaintances,only 11（10.51%）students learned it from school;the contact rates of eight kinds of high-risk behaviors were helping feeding 21.84%(62/206),holding lamb 16.50%(58/206),cleaning sheepfold 8.73%(26/206),vaccinating sheep 5.34%(17/206),milking 2.91%(6/206);cleaning apoblema 2.91%(6/206),slaughtering2.42%（5/206）and delivering lamb 1.94%(4/206)；there was no  significantdifference between the passing rate and the eight kinds of the basic protective behaviors except feeding (P>0.05).Conclusion The level of Brucellosis prevention knowledge is low in Western pastoral areas of Jilin province,and there was a high-risk of contact behavior with sheep and low basic protection rate.It indicates that the education about  prevention and  control of  Brucellosis in the students from breeding livestork’s familes   should be strengthened in order to prevent Brucellosis infection.

Objective To investigate the highly-recognized job evaluation factorsamong the doctors from tertiary general hospitals and to establish a doctor-oriented job evaluation factor system for general hospitals followed with the reliability and validity analyses.Methods A questionnaire about the highly-recognized 23 job evaluation factors was launched among 791 doctors by the stratified sampling from six tertiary general  hospitals in Beijing,and the factors were from a job evaluation model of  tertiary general hospitals.Initial factor solutions were obtained by the principal components analysis of all evaluation factors and the main factors whose eigenvalues were over one were extracted as evaluation dimensions.Factor loadings were attained through Varimax and the factor whose factor loading was lower than 0.5 was eliminated.A reliability analysis by calculating Cronbach αcoefficient and a validity analysis with structural equation modeling were sequentially conducted on the job evaluation factor system.Results A doctor-oriented job evaluation factor system for general hospitals including 22 factors subject to 3 dimensions was established while the factor of job relevance was removed.The internal consistency coefficients of the dimensions were greater than 0.8 according to the calculation of Cronbach α ,which showed a nice reliability. Several main indexes for evaluating the model fitting were all close to 0.9,which indicated a fair structural validity. Conclusion A doctor-oriented job evaluation factor system for general hospitals is established with fair reliability and validity,which could provide more references to the job evaluation of the doctors in  tertiary general hospitals.

Objective To explore the distribution and drug resistance of pathogen in hospitalized tumor patients and to provide reference for the clinical reasonable use of antibiotics and strengthening   the hospital infection control. Methods 6 500 clinical speciments were tested in hospitalized tumor patients from January  to December,2013. The drug susceptibilities were tested by automated microbiology system or Kirby-Bauer disk dilution method.  Drug susceptibility tests were evaluated according to CLSI standard 2012. WHONET5.6 software data were used to analyze the data. The clinical distribution and the resistance results of bacterial were analyzed retrospectively. Results A total of 2 093 strains of pathogens were isolated from 6 500 clinical speciments,among these strains，the gram-negative bacteria accounted for 55.23%,the gram-positive bacteria accounted for 11.08%,and the fungi accounted for 33.68%.  Klebsiella pneumoniae，Escherichia coli，Pseudomonas aeruginosa ranked the top three species of pathogens,accounting for 16.63%,9.60%,and 7.98%, respectively. Staphylococcus aureus,Candida albicans  ranked the first place of gram-positive bacteria and fungi, respectively.The antibiotic resistance of Escherichia coli was strong in Enterobacteriaceae,and its resistance rates to penicillins, cephalosporins,and  quinolones were more than 60%.Of the Staphylococcus,the methicillin-resistant Staphylococcus aureus(MRSA) accounted for 10.00% and the coagulase-negative Staphylococcus (MRCNS) accounted for 87.10%. There was no vancomycin resistant Staphylococcus aureus and coagulase-negative Staphylococcus detected.Enterobacteriaceae strains were found most sensitive to imipenem;gram-positive bacteria were found most sensitive to vancomycin.Conclusion The hospitalized tumor patients are susceptible to pathogens,and the  gram-negative bacteria are the predominant isolated pathogen. Etiology inspection and monitoring of antibiotics sensitivity provide experimental basis for clinical infection control and prevention.

Objective To explore the clinical variables associated with the shrinkage modes of primary breast tumor in women after neoadjuvant chemotherapy (NAC),and to develop a nomogram for predicting non-concentric shrinkage mode(NCSM).Methods Sixty-one women with pathologically proven solitary invasive ductal carcinoma(ⅡA-ⅢC) were recruited.Breast specimen was prepared with PMSS,and residual tumors were microscopically outlined,scanned and registered by Photoshop CS 5 software.The 3D model of residual tumors was reconstructed with 3D-DOCTOR 4.0 software to evaluate the shrinkage mode.17 factors such as age and body mass index and menopausal status were chosen as independent variables，and the clinic-pathologic shrinkage mode was considered as dependent variable.A Logistic regression model was used to construct the nomogram.Results Primary tumor stage,lymph node down-staging,PR and mammographic malignant calcification before NAC were independent predictors of clinic-pathologic shrinkage mode (β:1.538,OR:4.656,95%CI:1.414-15.328,P=0.011;β:1.555,OR:4.735,95%CI:1.082-20.722,P=0.039;β:-1.707,OR:0.181,95%CI:0.044-0.741,P=0.017;β:-1.405,OR:3.808,95%CI:0.06-0.998,P=0.048,respectively ).The nomogram predicting the risk of NCSM showed a good concordance index(0.869),and its conformity of mean absolute error was 0.039.Conclusion Based on the clinicopathological findings of primary breast tumor,a nomogram to predict shrinkage modes after NAC in breast carcinoma  patients is constructed.The statistical tool is helpful for individually selecting the patients who can be treated with BCT after NAC.