Abstract:Objective:To investigate the effects of oxidized low-density lipoprotein(ox-LDL) on the expression and activity of NAD-dependent deacetylase SIRT1 and its regulatory mechanisms on human adaptive immune function. Methods:Human primary peripheral blood monocytes were induced by human macrophage colony stimulating factor GM-CSF for 7 days in vitro. Well differentiated macrophages were treated with different concentrations(0~120 μg/ml) of ox-LDL for 48 h. The protein expression of SIRT1 was detected by Western blot. Real-time PCR was performed to examine the SIRT1 and HLA-DRα mRNA. Results:The protein expression of SIRT1 was decreased by ox-LDL in a concentration-dependent manner. The mRNA of SIRT1 treated by ox-LDL was significantly lower than that of the control group(P < 0.05).Resveratrol,which is the agonist of SIRT1,rescued the decreased expression of HLA-DRα induced by ox-LDL. Conclusion:The CIITA-dependent HLA-DRα promoter activity was decreased when treated with ox-LDL,which was induced by the change of mRNA and protein expression of SIRT1 in macrophage. Therefore,deacetylase SIRT1 may provide potential target for the treatment of atherosclerosis.

Abstract:Objective:To investigate the role of pyruvate kinase M2 (PKM2) on the invasion of liver cancer cell in vitro. Methods:We compared mRNA and protein expressions of PKM2 in hepatocellular carcinoma(HCC) tissues and adjacent normal tissues by real-time PCR and immunohistochemistry. Interfered and overexpressed PKM2 cell lines of HepG2 were constructed to analyze the role of PKM2 on invasion of liver cancer cells. Real-time PCR,immunohistochemistry and Western blot were performed to investigate the expression of HIF-1α and the phosphorylation of STAT3 in HepG2 cells. Results:The mRNA expressions of PKM2 and HIF-1α in the HCC tissues(106/115) were higher than those in adjacent normal tissues. Sample immunohistochemistry showed that the expressions of PKM2 and HIF-1α in HCC tissues were higher than those in adjacent normal tissues. In vitro experiment,interference of PKM2 inhibited the invasion of HepG2 liver cancer cell,and overexpression of PKM2 promoted the invasion of HepG2. The overexpression of PKM2 promoted the phosphorylation of signal transducer and activator of transcription 3(STAT3) and upregulated the expression of HIF-1α. Conclusion:PKM2 upregulates the expression of HIF-1α and promotes liver cancer cell invasion by promoting the phosphorylation of STAT3 and may represent a novel strategy for therapy of HCC.

Abstract:Objective:To explore the toxicity and dose-response relationship of 4-nm gold nanoparticles (AuNPs) on human lung adenocarcinoma cell line A549. Methods:Sodium citrate solution (the control group) and three concentrations (12.5,25.0 and 50.0 -滋g/ml) of 4-nm AuNPs solution were respectively added to the culture medium of A549 cells for 48 h incubation. The morphology and internalization of AuNPs in A549 cells were observed by transmission electron microscopy (TEM);cell viability of A549 was detected using cell proliferation and toxicity kit;cell cycle and apoptosis of A549 were analyzed by flow cytometry. Results:TEM showed that the shape and size of AuNPs particles were homogeneous and mainly distributed in small vesicles,cytoplasm,lysosomes or the perinuclear region in A549 cells. Compared to the control group,4-nm AuNPs significantly reduced the cell viability in a dose-response relationship (P < 0.05). The results of flow cytometry showed that AuNPs with higher concentration (25 and 50 -滋g/ml) significantly increased the apoptosis of A549 cells (P < 0.05),while no significant difference in cell cycle. Conclusion:AuNPs with size of 4 nm can significantly reduce cell viability and promote apoptosis in a dose-response relationship in A549,which provides a new strategy for the treatment of lung cancer.

Abstract:Objective:To study the effects of pigment epithelium derived factor (PEDF) on the expression of vascular endothelial growth factor(VEGF) in hypoxic human bronchial epithelial(HBE) cells,and to determine whether the expression of VEGF is associated with the expression of hypoxia-inducible factor (HIF)-1α in cultured cells. Methods:The cultured HBE cells between 3rd to 7th generations were used in the study,and 100 -滋mol/L cobalt (II) chloride hexahydrate (CoCl2) were used to simulate anaerobic condition. There were four groups in the study:control (group A),CoCl2 100 -滋mol/L(group B),CoCl2 100 -滋mol/L +PEDF 50 ng/ml (group C) and CoCl2 100 -滋mol/L +PEDF 200 ng/ml (group D). The expression of HIF-1α and VEGF mRNA were detected by real-time polymerase chain reaction (RT-PCR). The expression of VEGF in the different groups were examined by enzyme-linked immunosorbent assay(ELISA) and Western blot. Immunofluorescence (IF) was performed to test the expression of HIF-1α in cytoplasm and cytoblast. Results:①The level of VEGF mRNA was significantly elevated(8.56 ± 0.67)-fold in group B compared with that in group A (P < 0.01),which was decreased in group C and group D (P < 0.01) compared with that in group B. Compared with group A,the level of HIF-1α mRNA was elevated significantly in group B (P < 0.01). However,there were no significant differences of HIF-1α mRNA level among group B,C and D. ②The concentration of VEGF protein in cell cultural supernatant was higher in group B [(1 370.10 + 42.98)pg/ml,P < 0.01] than that in group A [(670.00 + 23.35)pg/ml]. The levels of VEGF in group C [(816.19 + 37.05)pg/ml] and group D [(646.47 + 22.70)pg/ml] were decreased compared with group B (P < 0.01). ③The level of VEGF protein detected by Western blot in group B was (3.99 ± 0.37)fold of that in group A (P < 0.01),and there were significant differences between group B and the other two groups (group C and D,P < 0.05). ④ IF showed that the level of HIF-1αprotein and the nuclear translocation of HIF-1α in group B were significantly increased compared with those in group A and D. Conclusion:PEDF could inhibit the overexpressed VEGF expression in hypoxic HBE cells,and this effect may be related to PEDF-regulated HIF-1α.

Abstract:Objective:To investigate the expression and effect of epidermal growth factor(EGF) on a rat model with colonic hypersensitivity induced by acetic acid. Methods:A total of 20 neonatal male rats were included in this study. Rat models with colonic hypersensitivity(n = 10) were established by intra-colonic infusion of 0.5% acetic acid(AA) at the age of 10~21 days,the controls(n = 10) received the same volume saline. Abdominal withdrawal reflex(AWR) score and external oblique muscle electromyography(EMG) activities were detected after colorectal expansion(CRD) stimulation to evaluate visceral sensitivity. The levels of EGF and 5-HT in plasma and intestinal tissues were measured by enzyme-linked immunosorbent assay(ELISA). The expression of serotonin transporter (SERT) was determined by Western blot to analyze the relationship between EGF levels and SERT expression in colon tissues. To further confirm the influence of EGF on SERT,rat intestinal epithelial cells (IEC-6) were stimulated at various concentrations of EGF (0,20,40 and 80 ng/ml) for 24 h to examine SERT protein expression. Results:Compared with the control rats,the AWR scores and EMG curve area during CRD in the rats with high colonic hypersensitivity were significantly increased (P < 0.05). HE staining and MPO level detection showed that no inflammation was found in both groups,which indicated that visceral hypersensitivity was successfully established. For visceral sensitized rats,the expression of SERT protein in colon were lower than that in the control group (0.711 ± 0.219 vs 0.980 ± 0.239,P < 0.01),and the levels of EGF in plasma[(2.639 ± 0.107)ng/ml vs(4.066 + 0.573)ng/ml,P < 0.05] and colonic tissues [(3.244 ± 0.135)ng/100 mg vs(3.582 + 0.197)ng/100 mg,P < 0.05] were also decreased compared with the controls. The concentrations of 5-HT in plasma and colonic tissues of model rats were significantly increased compared with those of the controls [(6.125 ± 0.534)ng/ml vs(3.540 ± 0.442)ng/ml,(5.527 ± 0.514)ng/100 mg vs(2.650 ± 0.495)ng/100 mg,respectively in plasma and colon,P < 0.05]. Furthermore,we analyzed the relationship between EGF and SERT level in colon tissues,and found that they were positively correlated (r = 0.820,P < 0.001). In vitro,IEC-6 cells were treated with EGF at different concentration (0,20,40 and 80 ng/ml) for 24 h. Compared with the concentration of 0 ng/ml,the relative expression of SERT was significantly increased (1.398 ± 0.091,1.725 ± 0.124 and 1.571 ± 0.088 at 20,40 and 80 ng/ml,respectively,P < 0.05). Conclusion:In visceral hypersensitivity rats,the EGF levels both in plasma and colonic tissue were decreased,but the 5-HT concentrations were increased. Moreover,the expression of SERT protein was positively correlated with EGF levels in colonic tissue. EGF can up-regulate the expression of SERT protein in IEC-6 cells in dose-dependent manner. These suggest that EGF may be involved in the formation of visceral hypersensitivity by affecting SERT expression.

Abstract:Objective:To explore the effect of different concentrations of D-glucose and different periods of time on autophagy in in vitro cultured podocytes. Methods:In vitro,cultured mouse podocyte clones (MPC5) were incubated with D-glucose at concentrations of 5.6,11,20 and 30 mmol/L for 6,12,24 and 36 h,respectively. The protein expressions of LC3 in podocytes were examined by Western blot and autophagosomes in podocytes were observed by transmission electron microscope. The changes of green fluorescent protein particles were observed after GFP-LC3 transfected podocytes by fluorescence microscopy. Results:①Compared with the 0 h group,the protein expression of LC3-II increased in a time-dependent manner and reach the highest at 24 h in podocytes exposed to 30 mmol/L D-glucose (P < 0.05). At the time of 24 h,the autophagosomes in podocytes and the green fluorescent protein particles in GFP-LC3 transfected podocytes were increased most prominent. ② Compared with the 5.6 mmol/L group,the protein expression of LC3-II was increased in a dose-dependent manner in podocytes exposed to D-glucose with increasing concentrations. In the concentration of 30 mmol/L,the expression of LC3-II was most prominent (P < 0.05). Conclusion:High glucose upregulates the protein expression of LC3-II in time- and dose-dependent manners and induces autophagy in cultured podocytes.

Abstract:Objective:To explore the expression of microRNA-21 (miR-21) in papillary thyroid carcinoma (PTC) and its effect on proliferation and apoptosis of K1 cell. Methods:Real-time quantitative PCR(qRT-PCR) was performed to detect and compare the expressions of miR-21 in 12 pairs of PTC tissues and adjacent normal tissues. K1 cell was respectively transfected with Anti-miR-21 and mimic-miR-21 and their corresponding negative control groups were constructed. The expression of miR-21 in K1 cells was identified by qRT - PCR. Cell proliferation after transfection with anti-miR-21 was analyzed using MTT assay. The cell apoptosis of K1 after transfection was analyzed by flow cytometry. Western blotting was performed to detect the expressions of proliferation and apoptosis-related proteins. Results:Compared with adjacent normal tissues,miR-21 in PTC tissues was significantly up-regulated (P < 0.05). Compared with the negative control (Anti-miR-NC) group,the miR-21 expression of K1 cells was significantly down-regulated after Anti-miR-21 transfection(P < 0.01). MTT assay showed that the inhibition of miR-21 expression significantly reduced the proliferation of K1 cells(P < 0.05). Flow cytometry showed that K1 apoptosis rate after Anti-miR-21 transfection was significantly increased to 19.5%,while the negative control (Anti-miR-NC) group was 9.4%. Western blotting showed that the expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated in the anti-miR-21 treated K1 cells. Conclusion:The expression of miR-21 is up-regulated in PTC. Down-regulated expression of miR-21 significantly inhibits proliferation while promotes the apoptosis of K1 cell.

Abstract:Objective:To investigate the effect of metformin on proliferation and apoptosis of human anaplastic thyroid carcinoma cells,and explore the underlying mechanisms. Methods:Human anaplastic thyroid cancer cell line SW1736 was treated with different doses of metformin. MTT assay was used to detect proliferation viability of cell line. Cell cycle progression and apoptosis were analyzed by flow cytometry. Results:Metformin significantly inhibited the growth of SW1736 cells in time- and dose-dependent manner with a maximal effect at 72 h. The results of flow cytometry showed that anaplastic cancer cell cycle was arrested to G0/G1 phase by metformin. Furthermore,metformin also enhanced apoptosis of SW1736 cells in a dose-dependent manner. Compared with the control group,addition of 20 mmol/L metformin for 48 h increased the percentage of apoptotic cells from 8.3% to 13.3% in SW1736 cells. Conclusion:Metformin can inhibit proliferation and cell cycle progression and induce apoptosis of thyroid carcinoma cell line. Therefore,it may be a potential therapeutic agent for the treatment of human anaplastic thyroid carcinoma.

Abstract:Objective:To study the effect of D-Rhamnose β hederin on the apoptosis of breast cancer cells and its effect on PI3K/AKT signaling pathway to explore its antineoplastic mechanisms. Methods:Breast cancer cell lines MCF-7 and MDA-MB-231 were treated with D-Rhamnose β hederin at different concentrations. Apoptosis rate of MCF-7 and MDA-MB-231 after treated with D-Rhamnose β hederin for 48 h were analyzed by AnnexinV/PI double staining of flow cytometry. Protein expression of PI3K/AKT signaling pathway related molecules after treated with D-Rhamnose β hederin were detected by Western blot. Results:D-Rhamnose β hederin with dosage of 20,30,40 -滋g/ml effectively induced cell apoptosis and the apoptosis rate was increased with the rising concentration. After treatment with 20 -滋g/ml D-Rhamnose β hederin for 48 h,the protein expressions of p-PI3K and p-AKT were decreased,but the total expressions of PI3K and AKT were not significantly changed. Futhermore,PI3K inhibitor LY294002 enhanced D-Rhamnose β hederin-induced apoptosis through inhibition of p-AKT. Conclusion:D-Rhamnose β hederin could effect on the expression of AKT phosphorylation and significantly induce the apoptpsis of breast cancer cells by inhibiting the phosphorylation of PI3K.

Abstract:Objective:To determine whether TGF-β1 induces the proliferation of Tenon capsule fibroblast (HTFs) and explored its mechanism. Methods:In vitro cultured 3~6 generations of HTFs were treated with various doses of TGF-β1 (0,0.5,1.0,2.0,5.0 and 10.0 ng/ml) for 24 h. Cell proliferation was detected by the cell counting kit-8 (CCK-8) assay and 5-ethynyl-2-deoxyuridine (EdU) incorporation assay. The expression of RhoE and its downstream cell cycle related protein cyclinD1 and proliferating cell nuclear antigen(PCNA) were detected by Western blot assay. The lentivirus was performed to establish the RhoE knockdown cell lines. Results:The proliferation of the TGF-β1-treated HTFs increased in a dose-dependent manner. Treatment with 5 ng/ml TGF-β1 caused a significant difference. EdU incorporation assay showed that the EdU labeling rate in the 5 ng/ml TGF-β1 group were higher than that in the control group. The average gray value of the expression of RhoE protein in HTFs in the 5 ng/ml TGF-β1 group was lower than that in the control group,while The average gray value of the expressions of cell proliferation protein cyclinD1 and PCNA were higher than those in the control group. After RhoE silenced in the cell,an increase of cell proliferation was detected by CCK-8 assay compared to the control group,flow cytometry showed that S period also increased and the gray value of cell proliferation protein cyclinD1 and PCNA were increased. Conclusion:TGF-β1 increases the expression of downstream cell cycle related protein cyclinD1 via downregulating RhoE,and induces the proliferation of Tenon capsule fibroblast.

Abstract:Objective:To determine the role of the parathyroid hormone (PTH) in fracture healing of mice. Methods:A total of 30 male mice at 8 weeks old (including 15 wild-type mice and 15 PTH gene knockout mice) were selected to establish right femur fracture model. At 7,14 and 21 days after the operation,the mice were sacrificed for measurement of bone mineral density (BMD) of the callus by micro-CT. To determine the role of PTH in fracture healing,the fracture models were assessed by histology,immunohistochemistry,X ray and micro-CT. Results:At 14 and 21 days after the operation,BMD in PTH gene knockout mice was significantly reduced compared to that in PTH wild-type mice (P < 0.05). X ray and micro-CT scanning showed that the fracture healing and the callus transformation and remodeling in PTH gene knockout mice were both delayed compared to those in wild-type mice. Conclusion:This study suggests that endogenous PTH promotes the fracture healing in mice.

Abstract:Objective:To explore whether the lack of endogenous PTH causes angiogenesis weakness by reducing the expression of VEGF during fracture healing and delays fracture healing. Methods:We constructed PTH knockouted mice fracture model,and observed fracture healing from X-ray film and detected bone anabolism and angiogenesis using immunohistochemistry. Results:VEGF expression was decreased in PTHKO mice. Immunohistochemical analysis showed that positive PEGAM was reduced. OCN activity was decreased and fracture healing was delayd. Conclusion:Lack of endogenous PTH may release VEGF by reducing mesenchymal stem cells or osteoblasts,reduce vascular fracture,inhibit osteoblastic activity,surpress endochondral ossification and lead to fracture healing delay eventually.

Abstract:Objective:To study the effect of endogenous parathyroid hormone (PTH) on osteoclasts (OCs) cultured in vitro. Methods:Sixteen PTH+/+ and sixteen PTH-/- mice with 8-week-old were used in this study. Bone marrow stem cells obtained from mouse femurs were isolated and cultured with macrophage colony-stimulating factor (M-CSF,30 ng/ml) and different concentration of receptor activator of NF-κB ligand (RANKL,50 ng/ml as low concentration group or 100 ng/ml as high concentration group) in culture dishes containing 10 ng/ml M-CSF for 24 h. The tartrate-resistant acid phosphatase (TRAP) staining was performed to observe and count cell morphology and TRAP-positive osteoclasts numbers at 40 or 100 magnified visual field after 6-day and 9-day culturing. Results:M-CSF and RANKL induced bone marrow stem cells to form TRAP-positive OCs. TRAP-positive OCs numbers in the same visual field decreased after 9-day culturing compared with that after 6-day culturing,but OCs volume after 9-day culturing became more larger and had more nuclei than after 6-day culturing. In the same concentration of RANKL,no changes were observed in the numbers of TRAP-positive OCs between the PTH+/+ groups and the PTH-/- groups. In contrast,either in the PTH+/+ or the PTH-/- groups,high concentration of RANKL (100 ng/ml) induce more TRAP-positive OCs. Conclusion:Endogenous PTH had no effect on the numbers of osteoclasts cultured in vitro in the presence of M-CSF and RANKL. High concentration of RANKL(100 ng/ml) increased the TRAP-positive osteoclast numbers compared with low concentration (50 ng/ml).

Abstract:Objective:To study the effects of trichostatin A(TSA) on the osteogenic differentiation of rat adipose-derived stem cells(ADSCs) in two-dimensional tissue culture plastic and three-dimensional nano-hydroxyapatite/collagen(nHAC) scaffold. Methods:ADSCs were cultured in two-dimensional tissue culture plastic (TCPT) and three-dimensional nHAC(nHACT) which contained 75 nmol/L TSA,and two groups without TSA(the TCP group and the nHAC group) were set as control. One and three days after ADSCs seeded onto the nHAC scaffold,the cells were characterized by field emission scanning electron microscope(FESEM). The alkaline phosphate(ALP) activity was detected to evaluate the early osteogentic differentiation at 3,5,7 d. Western blot was performed to detect the expression of Runx2,OPN and BMP2 proteins. Results:On nHAC,ADSCs were well adhered to and fully extended on the scaffold with TSA. ADSCs on TSA treated scaffold presented better. The data of ALP was continuously increased at 3,5,7 d. The highest expression of ALP occurred in nHACT had significant difference between groups(P < 0.05). Western blot demonstrated that Runx2,OPN and BMP2 protein in the nHACT groups increased more evidently than the other groups,with the same trends as ALP results. Conclusion:TSA could improve the early osteogentic differentiation of ADSCs,especially in the three-dimensional nHAC scaffold.

Abstract:Objective:This study aimed to explore the relationship between body mass index(BMI)and pulse wave velocity(PWV)in the community-dwelling subjects. Methods:A total of 3381 subjects older than 40 years of age from Gulou district,Nanjing were enrolled. Data of disease history,medication history,habits and etc. were collected by questionnaire. Height,weight,waist circumference and blood pressure were measured. Oral glucose tolerance test(OGTT)was performed. Blood lipid and PWV was assessed. Results:According to the multivariate regression,systolic blood pressure(SBP)was the independent risk factor of PWV(β = 0.478,P < 0.001). A weak and negative association was showed between PWV and BMI after adjustments for confounding factors by covariance analysis. The healthy obesity group had lower PWV than the unhealthy normal weight group(P < 0.01). Conclusion:BMI is not independently associated with PWV. The relationship between BMI and PWV is mainly explained by the strong positive association between BMI and BP.

Abstract:Objective:To investigate the difference of serum CETP activity between premenopausal and postmenopausal women with type 2 diabetes and its relevant metabolic factors. Methods:Fasting blood samples were collected from 113 subjects hospitalized in Department of Endocrinology,the First Affiliated Hospital of Nanjing Medical University from September 2011 to June 2012. All these subjects were female patients with type 2 diabetes before and after menopause. CETP activity and some biochemical markers of the subjects were measured. Results:Serum CETP activity of patients with type 2 diabetes was positively correlated with age,BMI,TG,FBG,HbA1c,and was negatively correlated with HDL-C. There was no obvious correlation between serum CETP activity and SP,BP,TC,LDL-C,Lp (a),fasting insulin levels,HOMA-IR,Cr,Urea,UA,ALT,AST,ALP and TBIL. Serum CETP activity of premenopausal women [(3.55 ± 0.24)-滋g/ml] was lower than that of postmenopausal women [(3.99 ± 0.28)-滋g/ml],and the difference was statistically significant(P < 0.01). Conclusion:CETP activity of premenopausal women is lower than that of postmenopausal women,and CETP activity is closely related with age and HDL-C levels.

Abstract:Objective:To investigate the relationship between serum 25-hydroxy vitamin D[25(OH)D]deficiency and thyroid autoi-mmunity among middle-aged and elderly population. Methods:A total of 2374 middle-aged and elderly patients were enrolled in this study. Serum 25(OH)D and thyroid autoantibody were determined. The levels of 25(OH)D and the prevalence of 25(OH)D deficiency were compared between the positive thyroid autoantibody group and the negative thyroid autoantibody group. Binary logistic regression was conducted to investigate the relationship between the levels of 25(OH)D and the prevalence of 25(OH)D deficiency and thyroid autoantibody. Results:Females had higher prevalence of 25(OH)D deficiency and positive thyroid autoantibody than males. The serum 25(OH)D level in the positive TPOAb group were significantly lower than those in the negative TPOAb group,and the prevalence of 25(OH)D deficiency(25(OH)D< 50 nmol/L) were significantly higher in the positive thyroid autoantibody group compared with those in the negative thyroid autoantibody group. Binary logistic regression analysis revealed that 25(OH)D< 50 nmol/L was a risk factor for positive TPOAb and positive TPOAb and/or TgAb after controlling for age,gender,body mass index (BMI),serum FT3 and FT4(OR = 1.313 and OR = 1.287,respectively). Conclusion:Patients with positive thyroid autoantibody have a high prevalence of 25(OH)D deficiency,and 25(OH)D< 50 nmol/L was a risk factor for positive thyroid autoantibody.

Abstract:Objective:To investigate the expression of Toll-like receptors 2(TLR2) in peripheral blood mononuclear cells(PBMCs)from patients with ovarian cancer,and its role in inducing the expression of IL-10. Methods:We collected PBMCs from 20 patients with ovarian cancer, 20 with benign diseases and 20 healthy females. Expression levels of TLR2 mRNA in PBMCs in the 3 groups were determined by real-time quantitative PCR and then compared. PBMCs were then stimulated with TLR1,TLR2 and TLR6 ligands. The expression and secretion levels of IL-10 in each troup were assessed by real-time PCR and FACS,respectively. Results:TLR2 were all expressed in PBMCs of the three groups, and the expression levels of TLR2 mRNA in PBMCs in patients with ovarian cancer were higher than the benign group and the healthy controls(both P < 0.05). There was no significant difference in TLR2 between the benign group and the healthy controls. After stimulated with HKLM(TLR2 ligand) for 24 h,IL-10 mRNA level was significantly higher in the ovarian cancer group compared to those in the benign controls and the healthy controls(Fold = 2.25,P < 0.05;Fold = 2.33,P < 0.05). There was no significant difference in IL-10 mRNA level between the benign controls and the healthy controls. Increased expression of IL-10 was also observed upon stimulation by FSL-1(TLR6 ligand) in ovarian cancer patients compared to those in the benign controls and the healthy controls(Fold = 1.95,P < 0.05;Fold = 2.16,P < 0.05). No signifiant difference was found between the benign controls and the healthy controls. Furthermore,there was an observable increased(P < 0.05) IL-10 secretion level upon stimulation by TLR2 ligand HKLM for 24 h in ovarian cancer patients(Medium = 150.46) compared to the benign controls(Medium= 38.86 pg/ml)and the healthy controls(Medium = 44.93 pg/ml),while no significant difference was identified between the benign group and the healthy controls. It was particularly noteworthy that PBMCs did not show significant up-regulation of IL-10 production in response to Pam3CSK4(TLR1 ligand) (M=46.70,61.53 and 31.11 pg/ml,respectively) and FSL-1 stimulation (M=20.20,31.12 and 35.45 pg/ml,respectively) in the 3 groups. Conclusion:TLR2 was highly expressed in PBMCs in ovarian cancer patients. IL-10 may be related to the immune evasion of ovarian cancer,which promotes the tumor progression by tumor immune escape.

Abstract:Objective:To study the proportion of smudge cells and prolymphocytic cells in lymphocytes from peripheral blood smear to predict prognosis value of chronic lymphocytic leukemia (CLL). Methods:We counted 200 lymphocyte in each patient’s routine blood smear,and calculated the proportion of smudge cells and prolymphocytic cells in lymphocytes. We detected the expression of ZAP70 or CD38 among CD5+CD19+ cells by flow cytometry. We analyzed the correlation of smudge cells,prolymphocytic cells,ZAP70+ cells and CD38+ cells. Results:In 53 patients with CLL,the median percentage of smudge cell was 24.5% (1%~65%),the median percentage of prolymphocytic cell was 5.3% (1%~51%). Smudge cells were negatively related to prolymphocytic cells (r = -0.317,P = 0.021). The proportion of smudge cells in 11 patients with ZAP70+ was significantly lower than that in patients with ZAP70- (P = -0.046),and the proportion of prolymphocytic cell was higher than that in patients with ZAP70- (P = 0.002),smudge cells proportion was negatively correlated with ZAP70- expression (r = -0.7818,P = 0.004). In 15 patients with CD38+,the proportion of smudge cells was lower than that in patients with CD38-(P = 0.024),and the proportion of prolymphocytic cells was higher than that in patients with CD38-(P < 0.001),the proportion of prolymphocytic cells was positively correlated with ZAP70 or CD38 expression (ZAP70:r = 0.685,P = 0.020;CD38:r = 0.575,P = 0.025). Conclusion:In CLL patients,smudge cells or prolymphocytic cells on peripheral blood smear and expression of ZAP70 and CD38 in peripheral blood were the same. The detection of smudge cells and prolymphocytic cells on peripheral blood smear is simple and easy,it can be used as a prognostic indicator of CLL patients in basic hospital which did not carry out flow cytometry.

Abstract:Objective:To explore advantages of high-throughput sequencing of cell-free DNA from maternal peripheral blood for detection of chromosomal aneuploidies in prenatal diagnosis. Methods:A total of 1 000 maternal peripheral blood samples from gravidas with more than 13 weeks of pregnancy were collected,and the cell-free DNA in the peripheral blood samples were detected using high-throughput sequencing to determine chromosomal aneuploidy. The cases of chromosomal aneuploidies and euploidies were validated by traditional chromosomal karyotype analysis and clinical follow-up of babies after the birth. Results:In the analyses of 1 000 cases,one case failed. The results of the remaining 999 cases indicated 18 cases of abnormalities,including 12 cases of trisomy 21,two cases of trisomy 18,one case of trisomy 13,one case of X monomers,one case of trisomy X,one case of XXY. From 12 cases of trisomy 21,there was one case of gemellary pregnancy,and the results of fetal karyotype analysis showed that one fetal was euploid and the other fetal was trisomy 21. One case of trisomy X was validated as euploid by karyotype analysis and the remaining detection results were consistent to those with karyotype analysis. The total positive detection rate of trisomy 21,trisomy 18,trisomy 13 and sex chromosome aneuploidy was 94.44%. No abnormal signs of chromosome aneuploidies (trisomy 21,trisomy 18 and trisomy 13) were found in follow-up of face and physical development of babies after the birth for all cases with negative results,and there were no false-negative cases. Conclusion:High-throughput sequencing of cell-free DNA from maternal peripheral blood has high accuracy for detecting fetal trisomy 21,trisomy 18 and trisomy 13. For sex chromosome aneuploidy detection,measurement indicators in this study should be further improved.

Abstract:Objective:To investigate the epidemiological characteristics of Coxsakievirus A16 (CVA16),and thus lay foundation for the comprehensive prevention and control of hand, foot and mouth diseases and development of CVA16 vaccines. Methods:Based on the active and passive surveillance systems built for enterovirus disease,we performed a surveillance of CVA16-associated diseases among 2 875 and 2 080 infants and young children from Donghai county and Baoying county,respectively,from March,2012 to February,2013. A unify investigation form was used to collect related information of CVA16-associated cases, and a series of throat and anal swabs were collected for real-time fluorescent quantitative PCR assay. Results:During the whole surveillance period, the incidence rates of CVA16-associated diseases were 16.73%(481/2 875) for Donghai County and 9.28%(193/2 080) for Baoying County. Only one peak of incidence, which occurred in May,2012,was observed in both counties. The addresses of reported CVA16-associated cases covered all towns in both two counties, but the incidence was different among different towns. All of the reported cases aged between 9 and 42 months. Although the sex ratios(male: female) were 1.59∶1 and 1.47∶1 for Donghai County and Baoying County,respectively,there was no significant difference of incidence between male and female in both two counties. Conclusion:From March,2012 to Feburary,2013,CVA16 was the major prevalent enterovirus in Donghai County and Baoying County in Jiangsu Province. The prevalence of CVA16 showed an obvious seasonal trend and regional difference.

Abstract:Objective:To investigate the value of two-dimensional speckle tracking imaging in assessing regional myocardial dysfunction in a mouse model of acute myocardial infarction. Methods:Twenty C57/B6 mice were randomly divided into two groups: acute myocardial infarction (MI) group (n = 10) and sham-operation (SO) group (n = 10). Echocardiography was performed three days after surgery. High frame rate two dimensional images were recorded in the left ventricular short axis views at the papillary muscle level and analysised at EchoPac workstation. Peak radical strain (PRS) and peak radical strain rate (PRSR) of each segment were measured at systolic period. Left ventricular internal diameter at diastole (LVIDd) and systole (LVIDs), left ventricular volume at diastole (LVVd) and systole (LVVs), ejection fraction (EF) and fractional shortening (FS) were measured with anatomical M-model echocardiography. Results:Compared with those of SO group, LVIDd, LVIDs, LVVd and LVVs of MI group increased significantly(P < 0.01) while FS and EF reduced(P < 0.01); PRS and PRSR decreased significantly in all segments of MI group (P < 0.01), compared with those of SO group; PRSR of anterosepetal, anterior and lateral segments in MI group decreased significantly than other segments(P < 0.05). Conclusion:Two-dimensional strain imaging could accurately quantify regional myocardial function in a mouse model of acute myocardial infarction.

Abstract:Objective:To evaluate the accuracy of endoscopic ultrasound ( EUS) in the preoperative TN staging for rectal cancer (RC). Methods:We searched CBM,CNKI,Wanfang and VIP databases from inception to Oct. 2013. Domestic articles related to the accuracy of EUS in the preoperative TN staging for RC were collected comprehensively. Related journals,conference proceedings,academic dissertations were also searched manually. Based on the principles and methods of Cochrane systematic reviews,data were searched,studied and extracted. Data analysis was conducted by Meta-Disc1.4 software. Results:Sixteen studies with 759 RC cases which met the inclusion criteria were included in this analysis. Pooling was conducted by both fixed and random effects models. Meta analysis showed that the accuracy of EUS in the preoperative T1-T4 staging for RC was high. Pooled sensitivity of EUS to diagnose T1-T4 staging were 87.0%,82.0%,87.0% and 76.0%,respectively. Pooled specificity of EUS to diagnose T1-T4 were 98.0%,93.0%,88.0% and 96.0%,respectively. The accuracy of EUS in the N staging was also high. To diagnose N staging,EUS had a pooled sensitivity of 76.0% and specificity of 81.0%. However,the accuracy of EUS in the preoperative T staging was superior than N staging. Conclusion:The results suggest that EUS combined with abdomen CT may be an effective tool for clinical preoperative diagnosis in TNM stage of RC,which plays an important role in clinical selection of surgical approach and treatment.