Abstract:Objective: To explore whether IRE1α can mediate unfolded protein response (UPR) during embryo development of xenopus laevis and rescue the developmental defects induced with tunicamycin. Methods: IRE1α mRNA or specific antisense oligonucleotids morpholino were microinjected to the 4 blastomeres of 4-cell stage embyos to overexpress or knockdown gene expression;the splicing of XBP1 was tested with RT-PCR to reflect the UPR;Tunicamycin was used to treat the embryos to induce the endoplasmic reticulum stress (ERS). Results: The splicing of XBP1 was increased when IRE1α was overexpressed while the splicing was decreased when IRE1α was knockdwon. Embryos developmental defects and the increase of XBP1 splicing were induced with tunicamycin while knockdown of XBP-1 partially rescued the defects and knockdown of IRE1α rescued the defects significantly and the increased XBP1 splicing. Conclusion: IRE1α can mediate UPR during xenopus development. Knockdown of IRE1α rescued the developmental defects induced with TM and partially by the IRE1α/XBP1 pathway.

Abstract:Objective: Produced curcumin’s precursors with toughen genes with ester key,to observe its different effect on growing cells between prostates’ and the normal diploid’s. Methods: Human prostatic cancer DU-145 cells were cultured and incubated with Boc-phenylalanine-curcumin (BPC) for 6-24 h. Cell viability was detected by MTT. Cell apoptosis rate was detected by flow cytometry. DU-145 cells were cultured and incubated with BPC for 1-7 d for cell grow curve detected by counting process. The people aortic smooth muscle cells(HASMC) acted as control. Results: After treatment with 10~40 μmol/L BPC for 6-24 h,the growth inhibition of DU-145 cells was 7.37%~66.87%(P < 0.05),in dose-time-dependent manners. Partial cancer cells presented morph-ological characters of apoptosis,with FSC-SSC scatter diagram. After treatment for 24 h,the cell apoptosis rate was 37.84%~47.12%(P < 0.05). Compared with curcumin of same concentrations,the control group HASMC apoptosis rate was 0.94%~4.23%(P < 0.05). Conclusion: The prodrug of curcumin could induce apoptosis of prostatic cancer DU-145 cells,and reduce the side effects on normal diploid cells. This provided a novel strategy for further exploration of tumor-targeted chemotherapeutic drugs.

Abstract:Objective: To construct tumor necrosis factor receptor-associated factor 6(TRAF6) and its small hair RNA (shRNA) expression vectors, and to assess their function on rat glomerular mesangial cell(GMC). Methods: The full-length TRAF6 CDS (HA tag) or three kinds of shRNAs targeting TRAF6 gene were synthesized and cloned into eukaryotic expression plasmid pcDNA3.1 and pGenesil-1/GFP,respectively. After confirmed by restriction endonuclease digestion and nucleotide sequencing,the recombinant plasmids were transfected into cultured rat GMC through NeonTM transfection system. Western blot assay was then used to confirm the expression of HA-TRAF6 and find out the optimal shRNA against TRAF6 gene. Results: It was verified by restriction endonuclease digestion and nucleotide sequencing that the constructed eukaryotic vectors were all correct. Western blot assay showed that the plasmids of pcDNA3.1-HA-TRAF6 could express HA-TRAF6 and the TRAF6 shRNA-1(shTRAF6-1) was able to silence the target gene effectively. Conclusion: The expression plasmid of TRAF6 gene and its shRNA were constructed successfully, which provides essential experimental tools for studying biological functions of TRAF6 gene in the future.

Abstract:Objective: To establish a GST pulldown assay for detection of Daam1 activity in eukaryotic cells. Methods: Prokaryotic expression vector containing fusion protein GST-RhoA was constructed. The correct plasmid was transfected into E. coli. BL21 stain and inducted the expression of the fusion protein by IPTG. SDS-PAGE and Western blotting were utilized to determine the corresponding recombinant proteins. The fusion protein with glutathione beads was pulled down to detect the activity of Daam1 in HEK293T cells. Results: GST-RhoA vector was successfully conducted and stably expressed fusion protein GST-RhoA. The activity of Daam1 was detected by GST pulldown assay,showing high activity of Daam1 in HEK293T cells. Conclusion: The activity of Daam1 was successfully detected by GST pulldown assay. This experimental scheme can be used for further studies of active Daam1 targeting in eukaryotic cells.

Abstract:Objective: To evaluate the effects of bFGF on proliferation and differentiation of rat cochlear sensory epithelial cells. Methods: Cochlear sensory epithelial cells of neonatal rats were isolated and cultured with different bFGF concentration. Bromodeoxyuridine (BrdU) labeling and immunohistochemical staining with calretinin were used to identify mitotic status and properties of culture cells. Results:① In 100 ng/ml and 200 ng/ml bFGF group,the BrdU positive cells were 36% and 35%,respectively,while in 1 ng/ml and 10 ng/ml bFGF group,the BrdU positive cells were only 3% and 7%. ② Percentage of calretinin positive cells varied from 0.5%,1.6%,7%,and 7.6% in 1 ng/ml,10 ng/ml,100 ng/ml,and 200 ng/ml bFGF,respectively. The percentage of the BrdU-positive and calretinin-positive cells in high bFGF concentration groups (100 ng/ml and 200 ng/ml) were significantly different compared with the control group. Conclusion: Proliferation and differentiation of cochlear auditory epithelial cells can be regulated by bFGF.

Abstract:Objective: To observe the effects of bufalin on overexpression of FN and CTGF in glomerular mesangial cells (MCs) of rat induced by high glucose,in order to explore the potential protective role of bufalin against diabetic nephropathy (DN). Methods: Cultured rat MCs were divided into 5 groups:control, high glucose(30 mmol/L),high glucose plus 5 × 10-9 mol/L bufalin,high glucose plus 5 × 10-8 mol/L bufalin,high glucose plus 5 × 10-7 mol/L bufalin. The cell toxicity of bufalin on MC were analyzed by Trypan blue method,expression of FN and CTGF was measured by RT-PCR and Western blot,respectively. Results: No cell toxicity was observed when bufalin was applied at the concentrations of 5 × 10-9 -5 × 10-7 mol/L. The expression of CTGF mRNA(P < 0.05) and protein(P < 0.05),FN mRNA level (P < 0.05) was significantly increased in the high glucose group compared with that of the control group. While the above-mentioned over-expression value induced by high glucose can be down-regulated by high,median,and low concentration bufalin in a concentration-dependent manner. Conclusion: Bufalin can inhibit the over-expression of FN caused by high glucose in MCs in a concentration-dependent maner,and the mechanism may be related to downregulating synthesis of CTGF,which indicates that bufalin deserves more investigation against DN.

Abstract:Objective: To study the indication of human recombinant luteinizing hormone(r-hLH) in GnRH antagonist protocol. Methods: Retrospective analysis. All the patients using GnRH antagonist protocol were enrolled in this study from Aug,2008 to Aug,2010. This study was divided into two parts. ① The first part enrolled 107 patients with ovarian hypofunction (FSH≥10U/L or antral follicle count≤6),80 of them without supplementation of r-hLH (group A),27 of them received r-hLH together with GnRH antagonist (group B). ② The second part enrolled 71 patients using r-hLH supplementation and GnRH agonist triggering,47 of them had normal ovary function (group C),and 24 of them had ovarian hypofunction (group D). The outcomes include mature oocytes rate,fertilization rate,cleavage rate,high-quality embryo rate,biomedical pregnancy rate,implantation rate and clinical pregnancy rate. Results: ① In group B,implantation rate(29.41% vs 19.46%) and clinical pregnancy rate (44.44% vs 31.25%) were higher than group A,but there were no statistical significance. IVF mature oocytes rate and E2/≥14mm follicle of group A was significantly higher than group B. ② In group D,IVF mature oocytes rate (100.00% vs 93.71%),IVF fertilization rate(79.52% vs 38.46%) and ICSI fertilization rate (84.85% vs 42.74%) were significantly higher than group C. In group D,implantation rate (28.89% vs 17.44%) and clinical pregnancy rate (41.67% vs 25.53%) were higher than group C,but there were no statistical significance. Conclusions: Recombinant human luteinizing hormone supplementation to GnRH antagonist protocol can improve ovary response and pregnancy outcome of ovarian hypofunction women. There has no reason for women with normal ovary function to add r-LH in GnRH antagonist protocol.

Abstract:Objective: To investigate the relationship between high sensitivity C-reactive protein(hsCRP) and metabolic syndrome in a general population. Methods: We randomly selected 609 subjects aged 18-74 years stratified by gender and age in Songqiao,a rural area of Jiangsu Province. A standardized questionnaire was used to collect information on medical history,smoking,alcohol intake and use of medications. Blood pressure was measured by mercury sphygmomanometer. Serum hsCRP concentration was measured using a high sensitivity BNprosec immunonephelometric assay. Metabolic syndrome was diagnosed according to the criterions of IDF (2004). Results: With increasing number of the metabolic syndrome components,serum hsCRP concentration of both genders was increased significantly (for men,adjusted P = 0.043;for women,adjusted P = 0.003). Logistic regression indicated that prevalence of metabolic syndrome was in proportion to increasing hsCRP levels(for total subjects,adjusted P < 0.01;for men,adjusted P = 0.002;for women,adjusted P = 0.001). In further partial correlation analysis,waist and serum triglycerides were found to be associated with hsCRP in total subjects and both genders,while systolic blood pressure and serum HDL cholesterol were related to hsCRP only in men. Conclusion: hsCRP was independently associated with both prevalence and individual components of metabolic syndrome in this study population.

Abstract:Objective: Enterovirus 71(EV71) and Coxsackievirus A16(Cox A16) are the main pathogens of the Hand,foot,and mouth disease(HFMD). In this study,we compared clinical manifestation and outcome of HFMD induced by EV71 and Cox A16. Methods: A total of 570 HFMD patients treated in Nanjing Children’s Hospital Affiliated with NJMU were enrolled in this study from January to June in 2011. Throat swabs were collected from these patients for detecting virus by real time PCR. Manifestations,outcomes for these patients were analyzed. Results: The average age of 350 patients with EV71 infection was (2.5 ± 1.7) years. The average age of 220 patients with CoxA16 infection was (2.3 ± 1.3) years. Patients with EV71 infection had a significantly longer hospitalization period(P < 0.001). The main manifestations were fever with a temperature above 39℃ and lasted longer (P < 0.01). EV71-induced myoclonic jerk,oral ulcer were significantly higher (P < 0.05). EV71-induced cough,febrile seizure were significantly lower (P < 0.05). However,central nervous system complications were significantly higher (P < 0.05). About 3.7% of EV71-infected children had neurologic sequela,and five patients with EV71 infection died of encephalitis and cardiopulmonary failure. All the patients with CoxA16 infection showed total recover. Conclusion: The main pathogen of HFMD was EV 71 in Nanjing region from January to June in 2011. Compared with those patients infected by Cox A16,the children with EV71 infection had serious clinical manifestations and worse outcomes.

Abstract:Objective: To determine the effects of various doses of alcohol in pecutaneous transluminal septal myocardial ablation (PTSMA) for obstructive hypertrophic cardiomyopathy (HOCM). Methods: A 1∶2 matched case-control study was performed in our hospital. Ten consecutive patients with HOCM were assigned to receive small dose of alcohol(≤2.0 ml),and they were compared with 20 cases receiving standard doses(> 2.0 ml). Early results of hemodynamic changes and left ventricular remodeling were determined invasively and by echocardiography before and after PTSMA. Results: In low-dose alcohol group,left ventricular outflow tract pressure gradient (LVOTPG) was (28.0 ± 8.8) mmHg immediately after PTSMA (P < 0.01),and (29.5 ± 6.6) mmHg at 3 months (P < 0.01),versus (69.9 ± 13.1) mmHg before the procedure. In standard group,LVOTPG was(27.5 ± 7.2) mmHg immediately and (28.1 ± 5.0) mmHg at 3 months after PTSMA(P < 0.01),vs(70.6 ± 13.4) mmHg before the procedure. The thickness of interventricular septum (IVS) was significantly decreased at 3 months post-PTSMA in both groups(P < 0.01). However,there were no significant differences in hemodynamic and morphologic changes between the 2 groups after PTSMA. No significant correlations were identified between the volume of ethanol injected and early hemodynamic, morphologic variables immediately or at 3 months after PTSMA either in low or standard dose of alcohol. Conclusion: Low-dose of alcohol was able to achieve the desirable degree of LVOTPG reduction and left ventricular remodeling in PTSMA.

Abstract:Objective: To investigate the effects of levosimendan(LS) combined with arginine vasopressin(AVP) on sepsis-induced lung injury(ALI) in swine. Methods: Fourteen female pigs were anesthetized and mechanically ventilated,and then peritonitis model was established. When mean arterial pressure (MAP) decreased to less than 60 mmHg,animals were randomly assigned to two groups: norepinephrine (NE) or LS (fixed dose of 0.4 -滋g/kg/min) + AVP (fixed dose of 0.57 mU/kg/min). If necessary,NE was titrated to maintain MAP between 65 and 75 mmHg in both groups. MAP,mean pulmonary artery pressure (MPAP),pulmonary capillary wedge pressure(PCWP),atrial oxygen saturation (SaO2),mixed venous saturation (SvO2),lactate concentration,hemoglobin concentration (Hb),cardiac output (CO),and plasma levels of TNF--琢,IL-6,IL-10,and NOax were determined at the following time points:before the establishment of sepsis model (baseline,T1),shock time(T2),and 6 h(T3) and 12 h(T4) after shock. Qs/Qt(venous admixture) was calculated at these time points. Time to the onset of septic shock point,total NE and fluid requirements were also recorded. Lung was removed for the determinations of pulmonary wet-to-dry ratio(W/D),myeloperoxidase activity (MPO),histological changes and apoptotic cells. Results: MAP,PCWP,time to the onset of septic shock point,total fluid requirements and Hb had no significant differences between the two groups(P > 0.05). Compared with NE group,MPAP,Qs/Qt,lactate concentrations,plasma levels of IL-6,and NOax at T4 and plasma levels of TNF--琢,HR and CO at T3 were significantly lower,while SaO2 and SvO2 were higher in LS+AVP group(P < 0.05). Total NE requirements,lung tissue levels of MPO activity,W/D ratio,total histological scores,and positive apoptotic cells were significantly lower in the LS+AVP group than those in NE group (P < 0.05). Conclusion: Combined LS with AVP improves gas exchange,decreases MPAP and lung inflammatory responses when compared with NE infusion alone in a porcine model of sepsis-induced ALI.

Abstract:Objective: To study the influence of hyperbilirubinemia upon newborns central nervous system by pass multiple-frequency auditory steady-state evoked responses analysis. Methods: Clinical data and examination results of multiple-frequency auditory steady-state evoked responses (ASSR) of 280 cases,blood total bilirubin consistency (TBC) higher than 220.6 μmol/L(12.9 mg/dl) month newborns,were retrospectively reviewed. Results: One hundred and ninty-five cases with TBC less than 310.0 μmol/L,had normal ASSR;18 cases with TBC higher than 520.0 μmol/L all took place encephalpathy of bilirubin,among them,16 cases of ASSR were heavy degree abnormal;67 cases with TBC less than 519.9 μmol/L or higher than 310.1 μmol/L,had no encephalopathy clinical performance,but ASSR appeard an abnormality,among them,63 cases (94.0%) were light-mid-degree abnormal,2 cases were heavy degree abnormal. The abnormality degree of ASSR is related with TBC,TBC and albumin ratio(TB/A)(P < 0.01),but wasn’t related to the constituent of pathogenesis of hyperbilirubinemia(P > 0.05). Conclusion: ASSR can suggest preclinical nervous poisonsing of hyperbilirubinemia in early stage,when TBC is higher than 307.8 μmol/L,ASSR should be included as a normal test.

Abstract:Objective: To explore the role of vitamin D in the pathogenesis of initial type2 diabetes mellitus (T2DM) by comparing the difference of blood serum 25-hydroxy-vitamin D[25(OH)D],clinical and biochemical indexes between initial T2DM and normal persons. Methods: Sixty-five patients with T2DM were enrolled and divided into T2DM A group and T2DM B group according to the level of serum 25 (OH)D, and 32 healthy people were selected as control. The levels of serum 25 (OH)D,clinical and biochemical indexes were compared between two groups, and the difference of relative indexes including waistline,BMI,SBP,DBP,FBS,HbA1C,PTH were compared. Results: The level of serum 25(OH)D in the case group was significantly lower than that in the control group. Serum 25 (OH)D was nagetively related to BMI,TG,Apo-A1 and PTH,and was positively related to HDL-C. There was no relationship between hypovitaminosis D and blood pressure,glucose or HbA1C. Conclusion: The level of serum 25(OH)D in the patients with initial T2DM was low,especially with obesity and abnormality in blood-fat. There was no relationship between serum 25(OH)D and blood pressure,glucose, calcium or glycosylated hemoglobin. The reason that T2DM patients were easily complicated by hypertension was possibly related to the rise of PTH, and the mechanism is expected to study.

Abstract:Objective: To explore the relationship between osteoprotegerin(OPG)/ receptor activator of nuclear factor-кB ligand (RANKL) system and the disease activity of ankylosing spondylitis and investigate serum osteoprotegerin and soluble RANKL (sRANKL) in evaluating the disease activity of ankylosing spondylitis. Methods: According to the criterion of Bath ankylosing spondylitis disease activity index and erythrocyte sedimentation rate, 22 ankylosing spondylitis cases with definite activity(A group) were matched with 22 ankylosing spondylitis cases with no evident activity (Q group). Serum OPG and sRANKL levels were determined by enzyme-linked immunosorbent assay. Results: Compared with Q group, a significant increasing level of serum sRANKL and sRANKL/OPG ratio was identified in A group, whereas the levels of serum OPG were comparable between them. Serum levels of sRANKL and sRANKL/OPG ratio were positively correlated with C reactive protein, erythrocyte sedimentation rate, Bath ankylosing spondylitis disease activity index and Bath ankylosing spondylitis functional index. Moreover, according to Youden index of ROC curve, the best cut-off point of serum sRANKL (> 4.76 pmol/l) and sRANKL/OPG ratio (> 0.73) for diagnosing disease activity emerged. The diagnostic sensitivity and specificity of cut-off point of serum sRANKL were 86.4% and 72.7%, whereas that of sRANKL/OPG ratio reached 90.9% and 86.4%, respectively. Conclusion: OPG/RANKL system may be associated with the activity of ankylosing spondylitis. Serum sRANKL/OPG ratio can be a promising clinical index in evaluating the activity of ankylosing spondylitis.

Abstract:Objective: To study the efficacy of concurrent chemoradiotherapy or radiotherapy alone in postoperative patient with N1 esophageal carcinoma. Methods: From October 2003 to October 2005,68 eligible patients with N1 esophageal carcinoma were randomized into radiotherapy alone group(RT,34 cases) and concurrent chemoradiotherapy group(CRT,34 cases). The schedules of radiotherapy in both groups were same,with 3-DCRT with conventional fractionation,and total dose is 54 Gy. The regiment of chemotherapy for the CRT group was consisted of DDP and 5-Fu for 2 cycles . It started on the first day of radiotherapy and repeated since the 29 day. Results: The overall survival rates at 1-,3-,and 5-year were 70.5%,38.2%,and 26.5% in the RT group,73.5%,47.1%,and 35.3% in the CRT group,respectively(P = 0.359);and the disease-free survival rates at 1-,3-,and 5-year were 58.8%,26.5%,and 14.7% in the RT group,64.7%,47.1%,and 26.5% in the CRT group respectively. The acute toxicity of chemoradiotherapy in the CRT group were higher than the RT group;later toxicity of two groups were similar. Conclusion: There was a trend of better survival in postoperative patient with N1 esophageal carcinoma treated with CRT than with the RT alone. However,the acute toxicity of chemoradiotherapy in the CRT group was higher than the RT group.

Abstract:Objective: Analysis of infection rate of mycoplasma pneumonia(MP) in Nanjing and surrounding areas are made in order to probe into epidemic characteristics of MP infection in this area. At the same time it offers help for prevention and treatment of the disease. Methods: Child outpatients with respiratory infection from Nanjing or surrounding areas in Nanjing Children’s Hospital Affiliated to Nanjing Medical University from January 2008 to December 2010 were subject to serum MP passive agglutination method to test MP-IgM antibody. Statistical analysis of MP-IgM antibody test result based on clinical data was made. Results: 31843 serum samples were tested during the three years. Out of them 13166 samples were positive,MP-IgM antibody with the positive rate being 41.35%;The positive rate in 2008 was the highest covering 45.08% of the total child patients infected with respiratory diseases in a year;Positive rate of MP-IgM antibody of children whose age was below or equal to 1 years old was 9.90%;From 1 to 3 years old was 32.48%;From 3 to 5 years old was 38.24%;Over 5 years old was 57.57%;Positive rate of male child patients was 36.08% and female child patients was 48.16%;On the other hand,Positive rate in the spring was 39.56%;In the summer was 36.51%;In the autumn was 42.11%;In the winter was 48.69%. Conclusion: MP is the main causative pathogen for respiratory infection of children in Nanjing area. MP infection goes on throughout the year with infection rate in the winter being the highest;Infection rate of infants is the lowest while that of preschool children is the highest;Infection rate of female child patients is higher than that of male child patients. The result of this study uncovers the epidemic characteristics of MP infection in Nanjing and surrounding areas,which is of significance for prevention and treatment of MP in this area.

Abstract:Objective: To investigate the epidemiologic characteristics of the deleted cases of syphilis and gonorrhea in Jiangsu province from 2006 to 2010, and to provide scientific references for the further report and control of these diseases. Methods: The deleted cases of syphilis and gonorrhea reported in China Information System for Disease Control and Prevention in Jiangsu province were collected and the data were analyzed. Results: The proportions of deleted cases of syphilis and gonorrhea in total reported syphilis and gonorrhea cases were increased;the experimental diagnostic cases were increased,while clinical diagnostic cases were decreased in deleted cases of syphilis and gonorrhea. Most deleted cases aged from 15 to 49 years old,and most cases were deleted by integrative hospitals;the majority of deleted cases were reported in the south of Jiangsu province. Conclusion: The deleted cases of syphilis and gonorrhea are increased in Jiangsu province,a series of strategies should be strengthened on cases report and prevention in sexually transmitted diseases (STDs) in order to improve the timely,integrated and accurate cases report of STDs.

Abstract:Objective: To study the effect of community methadone maintenance treatment among patients who had addicted to heroin in Lianyungang City. Methods: 480 patients who had entered the treatment center by the end of December 31,2009 were followed up for 1 year. Face-to-face investigation was conducted among 207 patients for baseline,6-month and 1-year evaluation with questionnaire by the doctor. The content is concerned to heroin abuse,present job status and social function. Results: The retention rate was 48.96% in this study. Along with the length of treatment,the proportion of both having a job and living with relatives increased in the two evaluation groups(P < 0.05). Social function and family function were recovered gradually. Heroin abuse behavior,injection and syringe-sharing behavior were statistically difference among the three groups(P < 0.05). With the exception of 2 HIV positive cases that had been infected before the program was initiated,none of the HIV negative patient had seroconverted to the HIV positive status. Conclusion: Services provided by Community-based methadone maintenance treatment was thought to reduce drug-use and HIV related high risk behaviors such as sharing syringes. Family function and social function were improved after treatment. In-depth counseling and support from the society were urgently required aimed at the individual cases.

Abstract:Objective: To assess the efficacy of glucocorticoids on the treatment of severe viral hepatitis. Methods: A comprehensive search for articles from 1995 to 2010 was conducted from databases including PubMed,EMbase,Science Direct,Wanfang,VIP,and CBM. The last search was performed on 10th October 2010. The quality of included studies was critically evaluated. Data were analysed using the Cochrane Collaboration’s RevMan 5 software. Results: A total of 683 articles were retrieved,but only 12 were finally included. Compare with the control group,meta-analyses showed that the mortality in glucocorticoids group reduced by 36%(RR=0.64,95%CI=0.42~0.98,P = 0.04),the effective rate in glucocorticoids group increased by 63% (RR=1.63,95%CI=1.35~1.98,P < 0.01). Both of them have significant differences. Conclusion: Compared with the control group,glucocorticoids tends to reduce mortality and improve liver function. However,it is necessary to do more well-desinged multi-center large-scale randomized controlled trials to confirm this.

Abstract:Objective: To observe the expression changes of toll-like receptor 2 and 4 (TLR2/4) in human gingival epithelial cells(HGECs) with and without lipopolysaccharides (LPS) treatment. Methods: TLR2/4 mRNA expressions were detected in HGECs by reverse transcription PCR. In addition,TLR2/4 mRNA and protein expressions were also determined in the cells stimulated with 1 μg/ml Porphyromonas gingivalis (P.gingivalis) LPS or 1 μg/ml Escherichia coli (E.coli) LPS by real-time PCR and FACS separately. Results: TLR2/4 mRNAs were expressed in HGECs from different individuals. Significant differences were found in TLR2 mRNA and protein expression in the cells with and without 1 μg/ml P.gingivalis LPS treatment(P < 0.05). In addition,significant differences were also found in TLR4 mRNA and protein expression in the cells with and without 1 μg/ml E.coli LPS treatment(P < 0.05). Conclusion: TLR2/4 were stably expressed in HGECs and their functions could be activated by LPS treatment,which may play a role in the development of periodontitis.

Abstract:Objective: To investigate the expression and clinical significance of proteinase activated receptor-1(PAR-1) in oral tongue squamous cell carcinoma(OTSCC). Methods: Real-time RT-PCR was used to characterize the expression of PAR-1 mRNA in 15 samples of OTSCC. The expression of PAR-1 protein in 109 tumor tissue samples including with peritumoral tissue from patients with OTSCC was detected with immunohistochemisty. Results: RT-PCR and immunohistochemisty showed that the expression of PAR-1 in OTSCC was significantly higher than that in the tumor-adjacent non-malignant tissues (P < 0.01). And the statistical analysis of clinicopathologic characteristics and immunohistochemistry showed that the high expression of PAR-1 protein in OTSCC was related to diameter of tumor and the later of stage grouping with TNM classification (P = 0.035 and P = 0.015,respectively). In contrast,no statistically significant correlation was found for gender and age. Kaplan-Meier survival and Cox regression analyses also showed that high PAR-1 expression and stage grouping with TNM were independent poor prognostic factors for OTSCC patients. Conclusion: The expression of PAR-1 protein was related with malignant level of OTSCC cancer. PAR-1 could be used as a mark of clinical prognosis and a potentially therapeutic target of OTSCC.

Abstract:Objective: To investigate the application value of dual-source computed tomography dual-energy angiography in carotid arterial. Methods: We retrospectively analyzed the images of 141 patients detected by dual- energy angiography,evaluated the quality of the images and used various reconstruction methods to analyze the pathological changes in carotid arterial. Results:High quality images were acquired in all of the 141 cases apart from 28 cases with no abnormalities and 9 cases showed normal variants of blood vessels,abnormalities of blood vessels were detected in 104 patients. 140 vessels with stenosis were detected in 99 patients with atherosclerotic plaque,including 6 patients with aneurysms,3 aneurysms and 2 arteriovenous malformation were detected in 5 patients without atherosclerotic plaque. Conclusion: As a non-invasive,cheap and convenient method,DE-CTA can remove bone in the neck and skull base,visualize stenosis,atherosclerotic plaque and aneurysms of carotidartery. DE-CTA possesses important effect to screen,diagnose,and track neck vascular diseases.

Abstract:Objective: To evaluate the effect of average heart rate,heart rate variability and selection of reconstruction phase on quality of coronary angiography image by 64-slice spiral computed tomography (CT). Methods: Fifty-one patients with suspected coronary artery disease or coronary atherosclerosis underwent CT coronary angiography using Siemens 64-slice spiral CT. Raw helical CT data were reconstructed at 0%~95% of the cardiac cycle with increments of 5% by Siemens Syngo MMWP VE23A reconstruction processing work station. The reconstruction included volume reconstruction,multi-planar reconstruction,maximum intensity projection and curved planar reconstruction. The 51 patients were divided into several groups according to the average heart rate and heart rate variability,and then select the optimal reconstruction phase and evaluate the quality of the image. Results: The optimal reconstruction phase of the coronary artery in < 65 beats per min group was 60%~75%. The optimal reconstruction phase of the coronary artery in 65-70 beats per min group was 65%~70%. The optimal reconstruction phase of the coronary artery in > 70 beats per min group was 30%~40%. There was a negative correlation between heart rate,heart rate variability and image quality score(P < 0.05). Conclusion: The appropriate control of heart rate and heart rate variability,and the correct selection of reconstruction phase could improve the image quality of coronary angiography.

Abstract:Objective: To research the blood perfusion characteristic of non-alcoholic fatty liver disease (NAFLD) using multisection dynamic CT. Methods: Dynamic 64-slice computed tomography scans of the liver were obtained in 72 individuals including 20 control subjects,52 patients with NAFLD from October 2009 to June 2010. The multisection dynamic CT perfusion series were performed by a multisection CT scanner (Philip Brilliance 64),120 kV,250 mAs. The data were processed on a Philip Brilliance 64 workstation using perfusion CT software package,and the mean HAP,PVP,TLP,and HPI were measured and statistically analyzed in 72 subjects. Results: Liver perfusion (TPL and PVP) in patients with NAFLD decreased (P < 0.05) and HPI increased significantly (P < 0.05). Conclusion: Hepatic perfusion parameters measured with CT were significantly altered in NAFLD. Dynamic 64-slice CT is potential in predicting prognosis of patients with NAFLD.

Abstract:Objective:To preliminarily evaluate the feasibility and potential values of whole-brain perfusion imaging performed with 256-slice CT to assess normal adult cerebral hemodynamics. Methods: Thirty-five normal results were selected from one hundred and fourteen patients who underwent brain CT unenhanced scan,CT perfusion imaging and CT angiography in head and neck for suspicion of ischemic cerebrovascular disease. The radiation dosage of CT perfusion imaging was recorded. Two senior neuroradiologic doctors independently analyzed the CT perfusion maps. Region of interest(ROI) was placed on bilateral temporal gray matter of two slices(the basal ganglia slice and body of lateral cerebral ventricle slice) supplied by middle cerebral artery,and the cerebral blood flow(CBF),cerebral blood volume(CBV),mean transiting time(MTT) and time to peak(TTP) values of ROI were automatically calculated by CT perfusion software. The mean values of perfusion parameters were compared by one-way ANOVA test. Results: The radiation dosage of thirty-five normal adults was 2.307 ± 0.008 mSv. There were no statistically significant differences between the two observers on the mean values of perfusion parameters(P > 0.05). The CBV and CBF values of the two observers between two slices had statistically differences(P < 0.05). Conclusion: With the lower radiation dosage and the stable and reproducible perfusion parameters,whole-brain perfusion imaging performed with 256-slice CT can be used to assess the entire cerebral hemodynamics.

Abstract:Objective: To find out the optimal separating conditions and wavelength program for measurement of 16 polycyclic aromatic hydrocarbons in house dust by high-performance liquid chromatography (HPLC) with fluorescence detector. Methods: Vacuum cleaner was used for gathering the house dust,and acetone was added and extraction was performed in a ultrasonic bath. The supernatant was concentrated and then added methyl cyanide to 0.1 ml. With methyl cyanide and water as mobile phase,PAHs were efficiently separated by gradient elution and detected by fluorescence detector and wavelength programming. Results: Except for acenaphthylene,15 PAHs had highly efficient separation within 28 min and showed good linearity with 0.5-50 ng/ml range. The relative standard deviations were 1.35%-5.89%,and the recoveries were 62.45%-110.36%. Conclusion: HPLC with fluorescence detection and wavelength programming could be used for detection of PAHs in house dust because of its quickness,simplicity,sensitivity and reproducibility.

Abstract:Objective: To obtain the purified human ovarian normal fibroblasts (NFs) preliminarily. Methods: The ovarian tissues were dissociated by 0.25% trypsin, after overgrowing on the bottom of the culture dish,the cells were adhered to the bottom of the dish repeatedly to depurate them. Then the cells were verified according to its morphology under the inverted-microscope and were further confined by a series of biological markers with immunocytochemistry. And its growth cure was observed. Results: The typical shape of ovarian NFs was fusiform, cords like,triangular,polyangular and irregular. Most of them contain Pseudopods-like structure. The NFs showed positive staining for vimentin,fibroblast specific protein-1, desmin and α-smooth muscle actin,negative staining for cytokeratin pan,cytokeratin 7, cytokeratin 20, CD31, CD90, MMP-2, MMP-3, MMP-9, CA125, CA19-9, CEA, fibroblast activation proteinα (FAP-α) and platelet-derived growth factor receptorα(PDGFR-α). Conclusion: Human ovarian NFs were cultured successfully,which provides a tool for further studies on ovarian cancer and carcinoma associated fibroblasts.

Abstract:Objective: To investigate the feasibility of establishmene of a reliable jugular vein - carotid artery transplantation rat model,and observe the pathological features of the vein grafts. Methods: The jugular vein - carotid artery transplantation model of SD rat was established by cuff technique. Grafts were harvested at 1 d,7 d,and 21 d,respectively,and underwent morphometric analysis by HE staining. The thickness of intimal media hyperplasia was measured by Digimizer image analysis software. Results: Eighteen vein graft animal models were established successfully. There were two rats died,and vein graft thrombosis at one animal. The overall success rate was 83.3%. The intimal media thickness of vein graft was (48.47 ± 11.28) μm after 1 d and increased to (66.05 ± 19.1) μm after 7 d,and increased to (300.44 ± 181.54) μm after 21 d (-字2 = 125.2,P < 0.01),and comparison between any two has a significant difference. Conclusion: The cuff technique of estabilish ment of vein graf model was simple and with high successful rate. The model can replicate the pathophysiology of human vein graft procedure exactly,consequently be widely used in experimental studies for human vein graft diseases.