Objective:To construct prokaryotic cell expression vector of human Eotaxin for expressing and examining its activity,and for constructing the N-terminal mutations of human Eotaxin.Methods: Extract the total RNA from peripheral monocyte,and Eotaxin gene coding region was obtained by RT-PCR.The fragment was sequenced and analysed after cloning into T vector,then clone to prokaryotic cell expression vector-PET30a~+ and expressed in BL21(DE?3).Results:The right cloning of human Eotaxin and its prokaryotic cell expression were obtained.Conclusion: The construction and prokaryotic cell expression of human Eotaxin lay a foundation for its more expressing and examining their activity,and for constructing the N-terminal mutations of human Eotaxin.

Objective To measure the changes of enamel diameter and depth after Er :YAG laser irradiation .Methods 80 enam-el samples were randomly divided into two supergroups according to irradiation time (10 s and 20 s) ,each supergroup was randomly divided into 5 groups according to different irradiation energy (0 .5W ,1 .0W ,1 .5W ,2 .0W and 2 .5W) ,separately underwent the Er :YAG laser irradiation with different time and different energy .Observed and measured the diameter and depth of each pit with a stereoscopic microscope after irradiation .Results After 10 seconds irradiation ,the differences of enamel diameter between illumi-nate power 0 .5W and 1 .0W ,0 .5W and 1 .5W ,0 .5W and 2 .0W ,0 .5W and 2 .5W were statistically significant(P<0 .05) ,the differences of enamel depth between two groups were statistically significant (P<0 .05) eliminating 1 .5W and 2 .0W .After 20 sec-onds irradiation ,the differences of enamel diameter between illuminate power 0 .5W and 1 .0W ,1 .0W and 2 .0W ,1 .0W and 2 .5W were statistically significant(P<0 .05) ,the differences of enamel depth were the same to 10 seconds irradiation .Conclusion The diameter and depth of enamel were changed under the irradiation of different time and energy ,which was helpful to clinical treat-ment of enamel caries .

Objective To construct prokaryotic cell expression vector of human Eotaxin mutants.Methods By point mutation,eight amino acid residues in the N-terminal(residues of 3-7)and N-loop(residue 14)regions of Eotaxin were individually mutated to methionine and residue 14 was delleted or methiomine was inserted after the residue 14,and then cloned respectively into prokaryotic cell expression vector-PET30a+.Results Eight N-terminal and N-loop mutants of human Eotaxin and their prokaryotic cell expression vector-PET30a+ were gained.Conclusion The successful construction of prokaryotic cell expression vector of human Eotaxin mutants lays a foundation for their expression and biological activity and for filtering antagonists of CCR3.

OBJECTIVE To investigate the clinical efficacy of using allogeneic amniotic membrane for myringoplasty in the treatment of traumatic perforation of tympanic membrane.METHODS 27 cases of traumatic perforation of tympanic membrane underwent myringoplasty with allogeneic amniotic membrane between July 2003 and February 2008 were analyzed retrospectively.The perforation healing rate and hearing improvement were studied.RESULTS All cases were followed-up for more than 6 months. The perforation healing rate was 96.3%and PTA gain was(12?3.7) dB.CONCLUSION The virtue of myringoplasty with allogeneic amniotic membrane is the plenty of service,convenient operation,little damage and good histocompatibility and effect.

Objective To study the correlation of the adiponectin receptor 1 gene rs1139646C/G and rs10920531C/A single nucleotide polymorphisms with Chinese Han women with polycystic ovary syndrome (PCOS).Methods For PCOS group (n =80) and control group (n =80),polymerase chain reaction (PCR) was used to amplify the upstream and downstream 250 bases of rsl139646C/G and rs10920531C/A,respectively.PCR products were directly sequenced.Results For rs1139646C/G loci sites between PCOS and control groups,all were the homozygous genotype CC,and no statistical significance was found for genotype frequency and allele frequency between two groups.For rs10920531C/A loci sites between PCOS and control groups,no statistical significance was found for genotype frequency and allele frequency between two groups (P =0.683,0.580),and no statistical significance was found for two loci with genetic linkage disequilibrium analysis (x2 =0.307,Fisher 's P =0.579610,Pearson' s P =0.579608).Conclusions Adiponectin receptor 1 gene rs1139646C/G and rs10920531C/A single nucleotide polymorphisms have nothing to do with Chinese Han PCOS women.Genetic linkage disequilibrium analysis has nothing to do with PCOS.The rs1139646C/G in Chinese Han population is relatively stable site,and no locus mutation is found.

Objective To study the expression of HGF , RAGE in placentas of patients with gestational diabetes mellitus and its relationship with maternal and neonatal prognosis .Methods The complete clinical datas of late pregnancy,GDM treated group 40 cases,GDM untreated group 23 cases,the control group 30 cases.Immunohisto-chemical method was used to detect HGF and RAGE in the placentas of three groups ,the maternal and neonatal prog-nosis were recorded.Results In GDM untreated group,GDM treated group and the controlled group placenta ,HGF and RAGE expressed.The expression of HGF was gradually increased ,and any comparative differences between two groups were statistically significant ( F =7.56, P <0.05 );the expression of RAGE in placentas was gradually declined ,the differences among the control group and GDM untreated ,GDM treated group were statistically significant (F=11.62,P<0.05).The rates of maternal and neonatal complications in the GDM untreated group were signifi-cantly higher than that of the GDM treated group and the control group (χ2 =7.34,13.21,9.75,6.67,15.23, 12.53,all P<0.05).Conclusion HGF and RAGE may participate in the occurrence and development of GDM and maternal and neonatal prognosis closely ,regular treatment can improve the prognosis of mothers and infants .

Objective To investigate the ability of single photon emission computed tomography (SPECT) and MRI in detecting skull-base invasion in nasopharyngeal carcinoma. Methods Sixty-one patients with nasopharyngeal carcinoma received whole body and skull-base tomography SPECT, and nasopharynx and skull-base MRI before radiotherapy. The results were double-blind compared and evaluated. Results The overall positive rates of skull-base invasion detected by SPECT and MRI were 51% and 46% (P=0.508). In paitents with headache, cranial nerve palsy or both, the rates were 83% and 86% (P=1.000) ,80% and 80% (P=1.000), 88% and 94% (P=1.000), respectively. In patients with T1+T2 and T3+T4lesions,the rates were 22% and 0(P=0.031) ,74% and 82% (P=0.250) ,repectively. In patients with N0+N1and N2+N3lesions,they were 50% and 48% (P=1.000) ,53% and 40% (P=0.500) ,respectively. The conformation rate between SPECT and MRI was 85%. Binary Logistic regression analysis showed that T stage was a risk factor for positive SPECT(χ2=4.23,P=0.040, OR=3.04). Headache tended to be a risk factor for both positive SPECT and positive MRI (χ2=3.13, P=0.077, OR=4.54;χ2=3.64,P=0.056,OR=12.00). Conclusions The detection sensitivity of SPECT in skull-base invasion in nasopharyngeal carcinoma is equivalent to that of MRI. The consistency between SPECT and MRI is good. Moreover, there is a good correlation between SPECT and symptoms, signs and stage. SPECT of skullbase tomography is necessary for patients with severe headache, negative CT and those who can not receive MRI. When SPECT result is positive,skull-base should be considered to be invaded and should be defined as gross tumor volume in radiotherapy planning.

A 54-year-old female patient with congenital heart disease had a persistent complete left bundle branch block three months after closure by an Amplatzer ventricular septal defect occluder. Nine months later, the patient suffered from chest distress, palpitation, and sweating at daily activities, and her 6-min walk distance decreased significantly (155 m). Her echocardiography showed increased left ventricular end-diastolic diameter with left ventricular ejection fraction of 37%. Her symptoms reduced significantly one week after received cardiac resynchronization therapy. She had no symptoms at daily activities, and her echo showed left ventricular ejection fraction of 46%and 53%. Moreover, left ventricular end-diastolic diameter decreased 6 and 10 months after cardiac resynchronization therapy, and 6-min walk dis-tance remarkably increased. This case demonstrated that persistent complete left bundle branch block for nine months after transcatheter closure with ventricular septal defect Amplatzer occluder could lead to left ventricular enlargement and a significant decrease in left ventricular systolic function. Cardiac resynchronization therapy decreased left ventricular end-diastolic diameter and increased left ventricular ejection fraction, thereby improving the patient’s heart functions.

OBJECTIVETo explore the role of toll-like receptor 4 (TLR4) on oxidized low-density/β₂-glycoprotein I/β₂-glycoprotein I (ox-LDL/β₂GPI/anti-β₂GPI) antibodies complex induced macrophage foam cell formation.

METHODSThe peritoneal macrophages were separated from TLR4 intact C3H/HeN mice and TLR4 defective C3H/HeJ mice. The cells were treated with ox-LDL, ox-LDL/β₂GPI, ox-LDL/anti-β₂GPI, anti-β₂GPI/β₂GPI, ox-LDL/β₂GPI/anti-β₂GPI, lipopolysaccharide (LPS) for 48 h and the foam cells were identified by Oil red O staining for intracellular lipids. The total cellular RNA and the protein lysates were collected. The levels of tissue factor (TF) mRNA in two groups were detected by real-time PCR (RT-PCR), and the expression of phosphorylated nuclear factor-κB (NF-κB) p65 was detected by Western blotting. Monocyte chemotactic protein-1 (MCP-1) secretion from peritoneal macrophages was determined by enzyme linked immunosorbent assay (ELISA) kits.

RESULTSCompared with C3H/HeJ mice, lipid droplets in the cytoplasm of peritoneal macrophages from C3H/HeN mice were significantly increased and phosphorylation-NF-κB expression was significantly upregulated after stimulating by ox-LDL/β₂GPI/anti-β₂GPI complex (P < 0.01). TF mRNA and MCP-1 expression were also upregulated post ox-LDL/β₂GPI/anti-β₂GPI complex stimulation [TF mRNA: 0.041 ± 0.023 vs. 0.005 ± 0.003; MCP-1: (6 200.2 ± 6.4) pg/ml vs. (803.3 ± 5.5) pg/ml, P < 0.01].

CONCLUSIONTLR4 can enhance ox-LDL/β₂GPI/anti-β₂GPI complex induced peritoneal macrophage foam cell formation via upregulating phosphorylation-NF-κB, TF and MCP-1 expression.

Animals ; Antigen-Antibody Complex ; immunology ; Atherosclerosis ; immunology ; metabolism ; Cells, Cultured ; Foam Cells ; immunology ; metabolism ; Lipoproteins, LDL ; immunology ; Macrophages, Peritoneal ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C3H ; NF-kappa B ; metabolism ; Thromboplastin ; metabolism ; Toll-Like Receptor 4 ; immunology ; metabolism ; beta 2-Glycoprotein I ; immunology

Objective To evaluate the left ventricular diastolic function of patients with normal left ventricular ejection fraction ( LVEF) by echocardiography and real‐time cardiac catheter measurement ,and improve the accuracy and reliability of echocardiographic diagnosis . Methods One hundred and twenty patients with know n or suspected coronary artery disease w ho underwent coronary angiography and left ventricular catheterization were prospectively selected from July 2017 to January 2018 in the Affiliated Hospital of Jiangsu University . According to the left ventricular end diastolic pressure ( LVEDP) real‐time measurement ,the patients were divided into groups of LVEDP ≤15 mm Hg ( 43 cases ) and LVEDP ＞ 15 mm Hg ( 77 cases) . General data were compared and the difference of echocardiographic parameters between the two groups were analyzed ,and the ROC curve of each echocardiographic parameter for diagnosing LVEDP was draw n . Results T he parameters including flow propagation velocity ( VP) ,the ratio of filling fraction of E and A ( E/A) ,early diastolic filling deceleration time ( DT ) ,the duration of mitral A ( A‐dur ,) mitral annulus velocity at the septal side ( e′sep) ,systolic pulmonary venous flow velocity ( PVs) ,diastolic pulmonary venous flow velocity ( PVd ) and PVs/PVd were used to the diagnosis of the increasing of LVEDP ,however their accuracies were low ( AUC between 0 .5～0 .7) . T he parameters including left atrial volume index ( LAVI ) , tricuspid regurgitation ( T Rmax ) ,mitral annulus velocity in lateral wall of left ventricle ( e′lat ) ,average e′,E/e′sep ,E/e′lat ,average E/e′,velocity of pulmonary vein atrial reversal ( PVa) ,pulmonary vein atrial reversal duration ( Pva‐dur) ,the difference between the duration of pulmonary venous A wave and mitral A wave( PvaD‐AD) were also used to the diagnosis of the increasing of LVEDP , but their accuracies were still poor ( AUC between 0 .7～0 .9 ) . According to the real‐time left ventricular pressure measurement and different parameters of echocardiography ,the multivariate regression equation :LVEDP＝ 0 .292 LAVI + 0 .35 PVa + 0 .04 T Rmax + 0 .075 ( PvaD‐AD ) －0 .109 PVs －6 .773 was put forward as a correction standard ,the accuracy of the diagnosis of LVEDP was significantly improved ( AUC ＝0 .922) . Conclusions T he assessment of left ventricular diastolic function needs to be performed comprehensively with multiple parameters . T he multiple regression equation can accurately evaluate left ventricular diastolic function in patients with normal LVEF .