Abstract:Objective:To construct a lentiviral vector containing HHV－6 U94 gene and observe the effect of U94 on proliferation and angiogenesis of vascular endothelial cells. Methods:The U94－6×His fragment was amplified from plasmid pSR2PH－U94 by PCR and cloned into the lentiviral vector (pLenti6.3－MCS－IRES2－EGFP). The recombinant plasmid was identified by restriction enzyme digestion and gene sequencing,and then cotransfected 293T cells with the auxiliary packaging components plasmids to obtain recombinant lentivirus containing U94－6×His gene. Endothelial cells EA.hy926 were infected with the recombinant lentivirus. After the Blasticidin screening and identification by RT－PCR and western blot,the resistant cell clones were selected. The proliferation and tube formation capacity of endothelial cells were examined by CCK－8 and tube formation assay respectively. Results:The U94 gene was successfully cloned to lentiviral vector pLenti－U94－IRES2－EGFP. The recombinant vector was packaged into 293T cells and the titer of the purified recombinant lentivirus was 2.35 × 107 TU/ml. The U94 stably expressing cell line EA.hy926－U94 was obtained after transfection with the recombinant lentivirus and Blasticidin screening. CCK－8 and tubule formation experimental results show that the proliferation and angiogenesis ability of EA.hy926－U94 were deteriorated significantly compared to the negative control cells EA.hy926－NC and normal cells EA.hy926. Conclusion:The pLenti－U94－IRES2－EGFP lentivitral expression vector was constructed and stably expressing human herpersvirus 6 U94 gene EA.hy926－U94 cell line was established successfully. U94 could inhibit endothelial cells proliferation and angiogenesis.

Abstract:Objective:To construct a prokaryotic expression vector pCold－Ⅱ－Rap and to optimize the expression conditions of ranpirnase (Rap) protein and to determine the protein activity. Methods:Recombinant pCold－Ⅱ－Rap vector was constructed,and positive clone was selected and transformed into E.coli.BL21. The recombinant protein expression was induced by IPTG. The Rap recombinant protein was purified,and detected by SDS－PAGE and Western blot. The RNase activity of Rap was analyzed by MTT. Results:The results showed that the purified ranpirnase was obtained by optimizing conditions of expression and purification, and degradation of tRNA could be detected by ranpirnase in vitro. The fusion protein can inhibit the proliferation of breast cancer cell MDA－MB－231,the inhibition ratios were 93.49% at the 4th day. Conclusion:The recombinant ranpirnase would be applied as a potential therapeutic drug,which could conjugated with a tumor specific antibody or antibody fragment.

Abstract:Objective:To investigate whether rat bone marrow－derived mesenchymal stem cells(BMSCs) could spontaneously differentiate into endothelial cells in normal culture condition. Methods:Primary rat BMSCs were separated and cultured to passage 7. The passage 3 and passage 7 BMSCs were prepared for this experiment. The expression of endothelial cell markers CD31,tyrosine kinase with immunoglobulin－like and EGF－like domains 2 (Tie－2) and von Willebrand factor (vWF) in passage 3 and passage 7 BMSCs were determined by Real－time PCR,and the ability of angiogenesis were determined by an in vitro angiogenesis kit (Matrigel). Futhermore,the expression of vWF protein in BMSCs were detected using immunofluorescence staining. Results:After normal cultured to passage 7,BMSCs partly formed an oval shape. Real－time PCR and immunofluorescence showed that the expression of endothelial cell markers were significantly higher in passage 7 BMSCs than in passage 3 BMSCs. In vitro angiogenesis experiments showed that passage 7 BMSCs could form a mesh structure in matrigel. Conclusion:BMSCs in normal culture condition changed their phenotypes,and could partially differentiate into endothelial cells spontaneously.

Abstract:Objective:To explore the osteogenic differentiation potential of de－differentiated mesenchymal stem cells (De－MSCs). Methods:Compared with MSCs,the proliferation capacity and osteogenic related－gene expression of De－MSCs were detected using CCK8 and qPCR assay. Meanwhile the osteogenic redifferentiation potential was measured by the ALP activity assay and the bone formation was confirmed by alizarin red staining. Results:De－MSCs could express stem cells markers,proliferation capacity was obviously higher than MSCs in osteogenic medium statistically. The expression levels of osteogenic related genes and ALP activities were remarkblely increased compared with MSCs. Red nodules were observed with alizarin red staining after De－MSCs were cultured in osteogenic medium for 28 days. Conclusion:De－MSCs retain some MSC traits,with a higher osteogenic differentiation efficiency than MSCs. De－MSCs could be an alternative potential seed cells in tissue engineering.

Abstract:Objective:To explore the role of Pioglitazone in the browning of both subcutaneous and visceral adipose tissue. Methods:Primary subcutaneous and visceral adipocytes were isolated from C57BL/5J mice and induced for differentiation. The primary cells were stimulated with 2.5 μmol/L Pioglitazone or vehicle throughout the whole induction period. Then the expression of either white fat or brown fat relative genes were observed by RT－PCR. Results:Pioglitazone could cause a significant increase of brown fat marker genes in the subcutaneous adipose tissue instead of the visceral adipose. Conclusion:Pioglitazone can promote a “white to brown” change in the subcutaneous adipose tissue instead of the visceral adipose.

Abstract:Objective:To observe the biological changes of human gastric cancer cell line SGC－7901, which was stably transfected with siRNA targeting osteopontin (OPN) and to study the molecular mechanism related to the biological changes. Methods:SGC－7901 cells were transfected with OPNsiRNA－pcDNATM6.2 and pcDNATM6.2 by lipofectamine 2000. Transfectants were selected and confirmed by Western blot and RT－PCR technique. The proliferation activity was detected by MTT assay and cell apoptosis was tested by flow cytometry. The migration of transfected cells was assayed by using transwell migration chambers. The expressions of OPN,CXCR4,MMP2 on mRNA and protein level were determined by RT－PCR and Western blot. Result:SGC－7901 cells were stably transfected with OPNsiRNA－pcDNATM6.2. MTT showed that the growth of OPNsiRNA transfected cells was slower than that of empty vecter transfected cells. OPNsiRNA gene transfection can increase the apoptosis. The migration of cells transfected with OPNsiRNA was suppressed compared with the cells transfected with empty vector,and the expressions of CXCR4 and MMP2 in SGC－7901 cells were significantly suppressed after OPN silenced by RNA interference. Conclusion:After OPN siRNA treatment,the cell proliferation and motility are suppressed,and apoptosis is enhanced;CXCR4 and MMP2 are OPN targeting genes. OPN may regulate the growth and metastasis of tumor cell via SDF－1/CXCR4 axis and MMP2 involved signaling pathway.

Abstract:Objective:To investigate the relationship between obesity and estrogen and their impact on uterus. Methods:Totally 48 rats were randomly divided into the following six groups:normal diet group(A),the high－fat diet group(B),uterectomy and normal diet group (C),uterectomy and high－fat diet group (D),estrogen supplement and normal diet group (E),estrogen supplement and high－fat diet group (F). The weight of the SD rats was tested weekly. The IPGTT experiment were taken after 14 weeks, and the experiment rats were executed after 16 weeks. The biochemical indicators were measured and the appearance of the uterus and the histological changes of different groups of the rats were compared. Results:Lack of estrogen increased the body weight and visceral fat content. TG,GLU,UA levels were negatively correlated with the estrogen level,while TG level was positively correlated with obesity. But there was no significant differences among these groups. The deficience of estrogen was associated with the insulin resistance,and the replacement of estrogen elevated insulin sensitivity and reduced blood glucose. The absence of estrogen led to analosis of uterus,and estrogen supplement significantly promoted the growth and development of uterus. Estrogen and obesity are both positively correlated with endometrial proliferation. Conclusion:Estrogen is closely correlated with obesity,and they both positively related to the growth and development of uterus and endometrial proliferation.

Abstract:Objective:To investigate the effect of gastric bypass on the expression of liver glucokinase(GCK) in non－obese Goto－Kakizaki (GK) type 2 diabetic rats. Methods:Twenty male GK rats and twenty male Wistar rats were randomly divided into four groups:GK operation group(GO group),GK sham operation group(GS group),Wistar operation group(WO group),Wistar sham operation group(WS group). There were 10 rats in each group. The fasting blood glucose and serum insulin levels were measured before operation and on the 1st,4th and 8th week after the operation. Liver tissues were harvested 8 weeks postoperatively and reverse transcriptase polymerase chain reaction was used to detect liver GCK mRNA expression after the operation. Results:The fasting blood glucose of GO group decreased as early as the 1st week after surgery,which decreased from preoperative (8.73 ± 1.30) mmol/L to (6.84 ± 0.89),(6.27 ± 0.93),(5.86 ± 0.57) mmol/L on the 1st,4th and 8th week postoperation (P < 0.05). The insulin level remained consistently higher in GO group with respect to GS group,but there was no significant change during the follow－up period. On the 8th week after surgery,the liver GCK mRNA level in GO group were significantly higher than GS group(P < 0.05). Conclusion:Gastric bypass can directly improve glucose metabolism in non－obese type 2 diabetic rats,the increased expression of liver tissue GCK mRNA might contribute to the hypoglycemic effect of the surgery.

Abstract:Objective:To study the anti－proliferative effect of vitamin D on the proliferation of airway smooth muscle cells(ASMCs) and prove the related mechanism of cyclin D1 and vitamin D in asthmatic rat. Methods:The ASMCs in male Sprague Dawley(SD) rats of control group and asthma group were cultured from primary generation. Cells between passage 3 and 5 were used in the study and were divided into control group,asthma group,asthma + 1 × 10-6 mol/L vitamin D group,asthma + 1 × 10-7 mol/L vitamin D group and asthma + 1 × 10-8 mol/L vitamin D group. The proliferation of ASMCs among each group was examined by MTT colorimetric assay and cell cycle analysis. The expression of cyclin D1 was detected by reverse transcriptase polymerase chain reaction(RT－PCR)and Western blot,respectively. Results:①The percentage of S(8.31 ± 0.42)% and G2/M(11.21 ± 1.43)% phase,the absorbance(A)(0.78 ± 0.042) value in ASMCs from asthma group were significantly increased compared with other groups;but the percentage of G0/G1[(80.01 ± 2.55)%] phase were significantly decreased,and compared to those(G0/G1,S and G2/M phase,A value) in ASMCs from control group[(90.15 ± 3.02)%,(5.41 ± 0.62)%,(4.38 ± 0.56)%,0.22 ± 0.04] and asthma + 1 × 10-6 mol/L vitamin D group[(89.08 ± 2.81)%,(5.58 ± 0.92)%,(4.61 ± 1.12)% and 0.38 ± 0.02,P < 0.001]. The percentage of G0/G1,S and G2/M phas,A value in ASMCs from asthma + 1 × 10-7 mol/L vitamin D group and asthma + 1 × 10-8 mol/L vitamin D group were compared with asthma group respectively(P < 0.01);②There were significant differences in the ratios of A value of cys D1 mRNA and expression of cyclin D1 protein in ASMCs from asthma group compared with control group and asthma + 1 × 10-6 mol/L vitamin D group respectively (P < 0.001) ;③And in the proliferation of ASMCs and the cyclin D1 mRNA and protein expression among different concentration of vitamin D groups,there were significant differences between the high and the low concentration(P < 0.001). Conclusion:ASMCs in asthmatic rats proliferate significantly and the expression of cyclin D1 increases obviously. Vitamin D can suppress the proliferation of ASMCs in asthmatic rats via regulating cyclin D1 expression.

Abstract:Objective:To investigate the effects of perifosine on the growth of human gastric carcinoma cell line SGC7901. Methods:The inhibitory effect of perifosine on the growth of human gastric cancer cell line SGC7901 was observed at different concentrations(0.125,0.250,0.500,0.750,1.500) -滋mol/L by sulforhodamine B(SRB)assay. Plate colony formation assay was performed to detect the colony formation rate. The expression levels of eIF4E and AEG－1 were determined by Real－time PCR and Western blot. Results:Perifosine inhibited the growth of gastric cancer cell line SGC7901 in dose－ and time－dependent manners. The cell colony formation rate was statistically decreased (P < 0.05) as compared with those in the control. The mRNA and protein levels of eIF4E and AEG－1 were reduced gradually dose－dependently. Conclusion:Perifosine,an AKT inhibitor,inhibits the growth and the colony formation rate of human gastric cancer cell via downregulating the expression of eIF4E and AEG－1.

Abstract:Objective:To investigate the relationship between the NAD (P)H oxidase p22phox C242T gene polymorphism and stroke. Methods:In this study,118 cases of cerebral hemorrhage,125 cases of cerebral infarction and 147 healthy controls were included,all cases are Han people in Shanghai region of China. Three groups had same baseline features of age,sexual proportion and BMI. Fasting glucose,and serum lipids were tested. The genotype and allele frequencies of the p22phox C242T polymorphism were determined by polymemse chain reaction－restriction fragment length polymorphism(PCR－RFLP) and gene sequencing technology. Results:The CT+TT genotype frequencies in the cecebral hemorrhage group,cerebral infarction group and control group were 9.3%,9.6% and 2.0%,respectively. T allele frequencies were 4.7%,5.2%,and 1.0%,respectively. The CT genotype and T allele frequency of the cerebral hemorrhage group and cerebral infarction group were significant higher than that of the control group (P < 0.05),one TT genotype was found in cerebral infarction group,and no TT genotype was found in cecebral hemorrhage group and control group,which was consistent with the results of gene sequencing. Binary logistic regression analysis was used to determine the correlation of CT genotype,cerebral hemorrhage (β = 1.712,OR = 5.537,95%CI:1.120~27.381,P = 0.036) and cerebral infarction(β = 1.432,OR = 4.187,95%CI:0.934~18.774,P = 0.061),respectively. Conclusion:These results suggest that the NAD(P)H oxidase p22phox C242T polymorphism was associated with cerebral hemorrhage and cerebral infarction,maybe serve as a risk factor. But compared to the traditional dangerous factors such as hypertension and alcohol,the effect of the C242T polymorphism on the cerebral infarction was not significant.

Abstract:Objective:To investigate whether any of the five single－nucleotide－polymorphisms (SNPs) of PON2 gene are associated with the susceptibility to occupational noise－induced deafness in Chinese Han population. Methods:A case－control study was conducted:188 cases whose average hearing threshold were no less than 40 dB in high frequency,and 200 controls in the same working position as the cases were matched in age,gender,the intensity and time of exposure to noise. Information on these subjects was gathered by questionnaire which was administered through face－to－face interviews by trained interviewers. Venous blood was gathered from all the subjects to extract out DNA which were genotyped by using the TaqMan－based allelic discrimination system. Results:rs7493(CG + GG) genotype,rs12026(CG + GG) genotype,rs7785846(CT + TT) genotype and rs7786401(GT + TT) genotype were the risk factors for occupational noise－induced deafness,adjusted OR(95%CI) were 3.54(2.18~5.75),3.38(2.08~5.49),3.30 (2.03~5.37) and 3.44 (2.11~5.62),respectively. When combined with noise exposure levels,the risk became much higher (OR increased). Conclusion:rs7493(CG + GG) genotype,rs12026(CG + GG) genotype,rs7785846(CT + TT) genotype and rs7786401(GT + TT) genotype of PON2 gene may be the risk factors for occupational noise－induced deafness in Chinese Han population. They may have interactions with the exposure level.

Abstract:Objective:To investigate the association of -347G→GA polymorphism located in the CDH1 gene promoter region with the expression of E－cadherin in sporadic colorectal cancer(CRC). Methods:The case－control study was designed,based on the population of colorectal cancer in China and healthy normal controls who have no tumor or inherited disease history. Polymerase chain reaction－restriction fragment length polymorphism(PCR－RFLP) was used to genotype the variant,and immunohistochemical staining was performed to measure the expression of E－cadherin in different allele cases among the CRC patients and normal controls. Results:There was no significant difference in the genotype frequencies of the polymorphism between normal controls and CRC patients(P > 0.05),there was no significant difference in E－cadherin expression for the GA－allele and G－allele in normal controls(P > 0.05),E－cadherin expression was lower for the GA－allele than for the G－allele in CRC patients (P < 0.05),which was no significant difference in normal controls (P > 0.05). Conclusion:The -347G→GA promoter polymorphism of CDH1 gene play no role in the etiology of CRC independently,but may be a prognostic factor during the progression of CRC.

Abstract:Objective:To investigate the association of plasma high sensitive C－reactive protein(hs－CRP) level and hemorrhagic stroke(HS). Methods:A case－control study(1∶2 matched) was conducted and 149 patients with acute cerebral hemorrhage from a county hospital and 298 age－ and sex－ matched controls from community－based population were selected in 2009. A unified questionnaire was managed to collect demographic information and history of disease. Blood pressure was measured and fasting glucose,plasma lipid and hs－CRP were detected. Cox model regression was applied to evaluate the association of hs－CRP level and hemorrhagic stroke. Results:The level of plasma hs－CRP in HS cases was significantly higher than that of controls (P < 0.001). The association of hs－CRP and HS was statistically significant and OR (95%CI) was 1.386 (1.230~1.563) for per 1 mg/L increasing (P < 0.001) after adjusted for histories of hypertension and diabetes,glucose and lipid. OR (95%CI) of hs－CRP elevation (≥3 mg/L) and HS was 6.599 (3.141~13.867) after adjusted for confounding factors (P < 0.001). Further stratification analysis by sex showed that men and women presented different associations of hs－CRP elevation and HS,and the ORs (95%CI) were 8.675 (3.206~23.469) and 3.836 (1.106~13.308) respectively. Conclusion:Our finding indicated that plasma hs－CRP elevation was significantly associated with increased risk of HS and the association was different between male and female.

Abstract:Objective:To assess Nanjing inhabitants’ knowledge,attitudes,and practice (KAP) with respect to epilepsy in Nanjing. At the same time advertised and popularized the knowledge about epilepsy to inhabitants. Methods:The door to door survey was used to interview inhabitants who give an informed consent and nor ever had an epilepsy seizure with knowledge,attitudes,and practice (KAP) questionnaire by stratified cluster sampling. Results:A total of 3 147 participants completed the questionnaire,there were 99.5% of participants had heard about epilepsy,among whom,31.9% knew at least one patient with epilepsy,and 41.7% had seen at least one epileptic seizure. 58.2% of participants thought epilepsy is an untreatable condition,5.0% of the respondents thought that epilepsy was contagious,17.2% thought it was a form of insanity. With respect to attitudes,12.3% and 84.1% would object their children to associate or marry with epilepsy patients. The independent determinants of attitudes were the belief that epilepsy is a form of insanity or contagious or is untreatable,and being married. Conclusion:The level of understanding and related knowledge of epilepsy affects attitudes to the patients with epilepsy,health education intervention about epilepsy should be carried out actively in this population,though Nanjing residents have a high awareness of epilepsy.

Abstract:Objective:To investigate the expressions of CXCR4,hTERT and microvascular density(MVD) in gliomas and discuss their relationships with the prognosis of glioma. Methods:Immunohistochemistry was used to detect the expression of CXCR4 and hTERT in 156 human gliomas,MVD was analyzed by VEGF labeling. Their relationships with survival time of patients were also analyzed. Results:① The positive rates of CXCR4 and hTERT in gliomas were 53.8%(84/156) and 50.6%(79/156), respectively. The positive rates of CXCR4 and hTERT in grade Ⅱ,grade Ⅲ and grade Ⅳ of gliomas were 30.8%(16/52),56.5%(39/69),82.9%(29/35),and 26.9%(14/52),53.6%(37/69),80.0%(28/35) respectively. A statistically significance of their expressions was observed among different grades(P < 0.01). ② The average of MVD in 156 gliomas was 44.17 ± 13.43. In grade Ⅱ,grade Ⅲ and grade Ⅳ of gliomas,MVD was 29.85 ± 4.34,50.12 ± 17.80 and 52.53 ± 18.16, respectively(P < 0.01). ③ The median survival time of patients with gradeⅡ,Ⅲ,Ⅳ of gliomas after operations was 30.2,11.7 and 9.8 months,respectively. The median survival time in cases with positive expression of CXCR4,hTERT or MVD≥44.17 were significantly shorter than that in cases with negative expression or MVD< 44.17(P < 0.01). Conclusion:The expressions of CXCR4,hTERT and MVD may be associated with invasive group and tumor angionenesis,their combined assay is helpful for evaluating prognosis.

Abstract:Objective:To study the classification of colorectal cancer tissue microvascular and the relationship between intratumoral microvascular density (MVD) and bevacizumab plus chemotherapy sensitivity in advanced colorectal cancer. Methods:Retrospective analysis was used to analyze 33 cases of advanced colorectal cancer treated with bevacizumab plus chemotherapy. Paraffin－embedded tumor samples were sectioned and stained immunohistochemically for the blood vessel markers CD34 and CD31 to characterize the intratumoral vasculature,then MVD were detected. Results:More blood vessels were stained by anti－CD31 antibody than anti－CD34 antibody. Two distinct types of microvascular were identified in colorectal cancer:differentiated (CD31+/CD34+) and undifferentiated (CD31+/CD34-). In the sensitive group,the number of undifferentiated－vessel MVD was 10.6 ± 3.1,significantly higher than 6.9 ± 3.7 in the insensitive group(P < 0.05). The median of each type of vessel was used as the cutoff value to divide the patients into high－ and low－MVD groups. Increased progression－free survival was seen in patients with high undifferentiated－vessel MVD(9.5 months vs. 3.0 months,P < 0.05). Conclusion:CD31+/CD34- MVD can be used as an important biology index to indicate the sensitivity to bevacizumab plus chemotherapy in metastatic colorectal cancer.

Abstract:Objective:To evaluate the effect of different load of hepatitis B virus on the serum lipids in chronic hepatitis B patients. Methods:The liver function parameters,the renal function parameters and the serum glucose of 412 hepatitis B patients were measured by automatic biochemical analyzer,the hepatitis B virus load were measured by quantitative PCR and the plasma prothrombin activity(PTA) were detected by automatic coagulometer. Results:The covariance analysis suggested that hepatitis B virus has no effect on the serum lipids (cholesterol,triglycerides,high－density lipoprotein cholesterol,low－density lipoprotein cholesterol) or the effect is negligible in the mild,moderate,severe chronic hepatitis B patients groups. While in the hepatic failure group,the hepatitis B virus has a significant effect on the serum lipids. The multiple linear regression analysis showed that the serum lipids in the hepatic failure group was mostly affected by the viral load (lgDNA), PTA,albumin. Conclusion:The different load of hepatitis B virus had different effect on the serum lipids in different stage of chronic hepatitis B patients. Through dynamic observation of lipid levels and viral load,we can understand the body immune status,as well as the role of hepatitis B virus in the body pathological immune response.

Abstract:Objective:To explore the efficacy and safety of anticoagulant therapy and local thrombolytics in patients with cerebral venous and sinus thrombosis(CVST). Methods:The clinical data of 24 patients with CVST were retrospectively analyzed. These patients were observed after discharge and followed up for 1~3 years. Results:In all the patients included,16 patients only anticoagulated with heparin,8 patients received anticoagulation with heparin and local thrombolytic treatment. At admittion,8 cases showed isolated intracranial hypertension,11 cases with focal deficit,2 cases with encephalopathy and 3 cases with coma. Both the anticoagulation and local thrombolytics were effective. At the end of the follow－up,17 cases (71%) were independent,3 cases (13%) were dependent and 4 cases(17%) died. Concomitant haemorrhageic infarcts related to CVST before treatment were not associated with a poor outcome. Independent predictors of poor outcome were mental disturbance,coma,multiple CVST while predictors of good outcome were thrombosis of single sinus and thrombolytic treatment followed by anticoagulant treatment. Conclusion:In our study,both the heparin therapy and local thrombolysis showed the safety and efficacy in patients with CVST. Local thrombolysis may be a therapeutic option for patients at high risk for a poor outcome despite heparin therapy,possibly in those with intracranial haemorrhages and coma.

Abstract:Objective:To investigate the clinical features of 84 female cases with primary bronchogenic carcinoma. Methods:A retrospective study of 84 female cases and 165 male patients with lung cancer that enrolled in Jiangsu province hospital was conducted. Results:During this study,the mean age of female patients was (59.2 ± 12.2) years. Compared with male patients,lower incidence of hemoptysis (P < 0.05),higher prevalence of peripheral lung cancer was seen in female patients,and the main pathological type was adenocarcinoma (P < 0.05). The incidence of lung adenocarcinoma in the cooking group is higher than the non－cooking group(χ2 = 10.2,P = 0.001). Conclusion:The female primary bronchogenic carcinoma is common,mainly middle－aged;the pathological type is mainly adenocarcinoma,closely relating to the cooking.

Abstract:Objective:To optimize the conditions for determination of the earthy and musty odors geosmin,2－methylisoborneol (2－MIB) in natural water by headspace solid－phase micro－extraction (SPME) coupled with gas chromatography/mass spectrometry and to establish a proposed method for determining the typical odor compounds in water. Methods:By applying the orthogonal experiment design L32(49) to arrange the determination conditions,several parameters of the extraction and desorption were studied and optimized (such as extraction temperature,desorption temperature,extraction time,ionic strength and rotate－speed),and some other parameters (such as correlation coefficient,limits of detection,RSD and recovery) were applied to evaluate effect of this method. Results:The optimum temperature of extraction and desorption were 65℃ and 260℃,respectively;time of extraction and desorption were 40 min and 7 min,respectively;ionic strength,30%(W/V);rotate－speed,600 rpm. Under the optimized experimental conditions,limits of detection(S/N=3) were 0.04 and 0.42 ng/L for geosmin and MIB,respectively. Calculated calibration curves gave high levels of linearity with correlation coefficient values of 0.9999 for both of geosmin and MIB. Finally,the proposed method was applied to the analysis of seven standard water samples,which were previously analyzed and confirmed free of target analytes. The RSD were 2.75%~3.80% and 4.35%~7.6% for geosmin and 2－MIB,respectively,and the recovery values were 91%~107% and 91%~104% for geosmin and MIB,respectively. Conclusion:The method can be successfully applied to analyze earthy and musty odors in natural water.

Abstract:Objective:To explore the method of computer discriminant on adenocarcinoma cells (ACC) and reactive mesothelial cells (RMC) amd establish the foundation on computer diagnosis for ACC in hydrothorax. Methods:Totally764 ACC of 44 cases and 709 RMC of 39 cases were collected from hydrothorax smears. Technique of computer image analysis was applied to test chromatics and morphological structure parameters of above cells. Approximately 80% cells were randomly selected as train group,and residual 20% cells as test group,the stepwise discriminant analysis was used in train group to establish the discriminant function,and the discriminant effects of function in test group were evaluated. Results:Stepwise discriminant showed that RP,BP,Rn,Bn,dmaj,c-dmin,c-An-Dn-Cn entered discriminant function,and its coincidence rate was 84.5%. The discriminant effect was significantly better than that in functions individually based on chromatics and morphological structure parameters(P < 0.05). Conclusion:The function combined with chromatics and morphological structure parameters has good discriminant effect,and it provides the analysis path of computer quantitative diagnosis for ACC in hydrothorax.

Abstract:Objective:To construct a three－dimensional finite element(FE) model of non－constrained artificial disc (NAD) replacement and lumbar vertebrae L4~L5 segment as the standard digital model for further biomechanical research. Methods:The NAD FE model was designed by using Auto CAD and it was combined with lumbar segment FE model in aforementioned part one by Abaqus software. The L4~L5 model was validated by Force 1,a 400 N axial load which was made up of four 100 N stress force,then Force 2 which was composed of 400 N axial load and 10 N-m extension bending moment. Results:L4~L5 segment model was consisted of 33,976 nodes and 162,858 elements. Centrum was consisted of 161,679 tetrahedron elements. Endplate was consisted of 1,053 hexahedral elements. Ligament,articular capsule and fiber were consisted of 126 Link elements. The NAD model was consisted of 11,583 nodes and 44,927 elements. Endplate and nucleus pulposus was consisted of 1,053 and 3,724 hexahedral elements,respectively. The results of load test showed that the model of NAD and L4~L5 segment had good effect of visual and mathematics. Conclusion:Based on mimics software and CAD methods,the 3D FE model of lumbar motion segment and artificial disc can be constructed automatically with excellent simulation results. Mimics can provide precise model through calculation based on threshold and parameterization modeling. The implanted L4~L5 segment can be used for further biomechanical research of lumbar spine.

Abstract:Objective:To evaluate the effects of surface treatments on bond strength of pressed and build－up veneering porcelain to zirconia. Methods:CAD/CAM Y－TZP blocks were fabricated and classed into 6 groups. Groups A1 and B1 received no treatment (as the control group),groups A2 and B2 received dye－penetrating technique,groups A3 and B3 were tribochemically silica coated. Then,groups A1 to A3 and groups B1 to B3 were veneered using build－up technique or lost－wax press technique respectively. The double－layered specimens were subjected to shear bond strength(SBS) testing. Data were statistically analyzed using one－way ANOVA and LSD test (α=0.01). The treated Y－TZP surface and the typical fractured Y－TZP surfaces were examined using a scanning electron microscope. Results:No matter which methods were used for treating Y－TZP surface,there was no statistically significant in the mean SBS values between the two veneering techniques. When using build－up veneering technique,tribochemically silica coating provided the highest SBS,while no statistically difference was found among the surface treatments of Y－TZP blocks for the lost－wax press technique groups. Conclusion:Dyeing and veneering techniques had no influence on bond strength of zirconia to veneering porcelain,while silica coating significantly improved the bond strength for the build－up veneering technique.

Abstract:Objective:To evaluate the effects of self－made all ceramic pin on the micro tensile bond strength of a rensin cement to dentin. Methods:Forty extracted molars were selected and randomly divided into four groups,group A,B and C were prepared with an internal pin channel,then the all ceramic pins,self－threading retentive pins and resin composite pins were inserted into dentin respectively,group D was prepared with no internal retentive features. All subjuects were filled with composite resin. For the microtensile nontrimming technique,10 beam－shaped specimens per group were obtained. Each specimen was loaded in tension until failure and the fracture mode present were observed under stero－microscope. All the results were analyzed statistically. Results:The micro tensile bond strength of group A and B was statistically higher than that of group C and D(P < 0.05),while there were no significant difference between group A and B,or between group C and D(P > 0.05). The breakage modes were observed mainly between the dentin and adhesive interface in group A,C and D,while the dentin cohesive fracture was appeared mainly in group B. Conclusion:IPS e.max Press ceramic pin can significantly enhance micro tensile bond strength of a rensin cement to dentin and is considered as the novel and useful dentin pin.