Objective To observe the expression changes of peripheral endothelial progenitor cells (EPCs) and Ang-1/Tie2 in patients with pulmonary arterial hypertension (PAH).Methods From Jun 2011 to Dec 2012,45 patients with PAH charged in Affiliated Hospital of Hangzhou Normal University were divided into 3 groups according to mean pulmonary arterial blood pressure (15 per group):mild(Group L),moderate(Group M),and severe(Group S),with another 15 normal people as control group(Group C).The EPCs were isolated from peripheral blood of every patient,number counting using fluorescence activated cell sorter (FACS),function test using cell culture in vitro.Expression of Ang-1 and Tie2 in the peripheral EPCs were measured by RT-PCR or Western-blot.Non normal data was analyzed by non parametric statistical test.Results The statistical discrepancy existed among Group L,M,S and the control in the number of EPCs [32.0 (27.0,37.0),26.0 (19.0,31.0),24.0 (22.0,26.0) vs 40.0 (37.0,51.0),P ＜ 0.05].The ability of migration [32.1 (26.5,37.5),26.8 (22.4,35.4),21.0 (17.8,34.0) vs 39.0 (33.3,42.4),P＜0.05] and adhesion of the EPCs [57.1(50.9,61.8),51.8(45.2,58.7),46.0 (37.2,55.1) vs 64.1 (56.2,75.0),P ＜ 0.05] among study groups and control group was different in statistic,the same with the proliferation activity of EPCs in different groups [0.6 (0.5,0.7),0.5 (0.4,0.6),0.4(0.3,0.5) vs 0.7(0.6,0.8),P ＜0.05].The mRNA expression of Ang-1 and Tie2 in Group M & S were significantly reduced compared with control [4.33 (2.49,4.62) and 2.89 (2.39,3.44) vs 5.31(3.78,6.22),P＜0.05],Tie2 mRNA[1.32(1.23,1.34)and 1.23(1.08,1.42)vs 1.49(1.25,1.66),P ＜ 0.05],and the protein expression of the phosphorylated Tie 2 in Group M &S were decreased [0.16 (0.15,0.24) and 0.12 (0.08,0.18) vs 0.22 (0.19,0.28),P ＜ 0.05].No significant difference of Ang-1 and Tie2 expression was observed between Group L and control [5.42 (4.72,5.95),1.54 (1.43,1.66) and0.23(0.19,0.33),P=0.674,0.867 and 0.674].Conelusion With the severity of PAH,the number and function of circulating EPCs decreased,as consistent with Ang-1 and Tie2 expression changes,suggesting that function decrease of EPCs in patients with PAH may be associated with the decrease of Ang-1/Tie2 expression.

Objective To determine the expression and clinical value of neuropilin-1 (NRP-1) in hepatocellular carcinoma (HCC).Methods One hundred and fifty-one cases of HCC tissues and 89 cases of healthy liver tissues were chosen to compare the expression of NRP-1 by immunohistochemistry.Then the relationships between different clinical factors and the expression of NRP-1 were analyzed by univariate and multivariate statistical analysis.Moreover,the survival rates were compared by survival analysis between different expressions of NRP-1 in HCC patients.Results Eleven cases were lost to follow-up or died for non HCC disease,and the effective cases in the final study were 140 cases.The positive expression rates of NRP-1 in HCC and normal liver tissues were 65.00％ and 35.96％ respectively,and the difference was statistically significant (x2 =18.843,P ＜0.001).According to the expression level of NRP-1,140 patients with HCC were divided into negative expression group (n =49) and positive expression group (n =91).Univariate analysis showed that the expression of NRP-1 in HCC was correlated with tumor number (x2 =8.025,P =0.005),TNM stage (x2 =26.467,P ＜ 0.001),differentiation degree (x2 =15.296,P ＜ 0.001),portal vein invasion (x2 =9.054,P =0.003) and hepatic vein invasion (x2 =5.928,P =0.015).Multivariate statistical analysis showed that TNM stage (OR =1.392,95％ CI:1.121-1.730,x2 =8.950,P =0.003),differentiation degree (OR =1.469,95％ CI:1.102-1.958,x2 =6.862,P =0.009),portal vein invasion (OR =1.829,95％ CI:1.157-2.893,x2 =6.665,P =0.010) and hepatic vein invasion (OR =2.161,95％ CI:1.172-3.987,x2 =6.084,P =0.014) were important factors for NRP-1 expression.The median survival time of NRP-1 negative HCC patients was significantly longer than that of positive group (44 months vs.23 months),and the difference was statistically significant (x2 =21.922,P ＜0.001).Conclusion NRP-1 is over-expression in HCC tissue and related to the malignant progress of HCC,and this suggests poor prognosis in patients with HCC.

BACKGROUND:There is a very close relationship between osteoporosis and periodontal disease in postmenopausal women, but the mechanism remains unclear. OBJECTIVE:To investigate the expression of nuclear factor-κB in the alveolar bone andinterleukin-17 in the serum and gingiva in the mouse model of osteoporosis caused by ovariectomy. METHODS:Female mice aged 3 months were randomly divided into ovariectomy and sham operation groups. At 6 months after surgery, the mouse models were evaluated histologically on the submandibular bone and thigh bone stained with hematoxylin and eosin. In the submandibular bone, the expression levels of OCN and Runx2 were detected by RT-PCR, and the expression level of nuclear factor-κB was detected by immunohistochemical staining and western blot assay. Besides, the expression level of interleukin-17 in the serum and gingival homogenate was evaluated using Cytometric Beads Array. RESULTS AND CONCLUSION:The thigh bone in the ovariectomy group revealed the thin cortical bone, enlarged marrow cavity, and increased resorption lacunae, as well as fewer, thinner trabeculae with lower density and irregular structure. Compared with the sham operation group, the expression levels of OCN and Runx2 in the alveolar bone were decreased in the ovariectomy group. The activation of nuclear factor-κB (P65)appeared with P65 positive expression in the submandibular bone in the ovariectomy group, and the relative expression level was higher than that in the sham operation group. The serum level of interleukin-17 in the ovariectomy group was higher than that in the sham operation group, but the level in the gingival tissue showed no significant difference between the two groups. These results indicate that estrogen deficiency after ovariectomy can activate nuclear factor-κB signal pathway to play a role in periodontal osteolysis. However interleukin-17 in the local periodontal tissue may not be a key cytokine to damage the periodontal tissue.

Objective To investigate the risk of hepatitis B virus (HBV) infection in the children born to HBV‐infected parent (s) after in vitro fertilization (IVF) .Methods From January 2008 to December 2013 , in the reproductive medicine center of the Third Affiliated Hospital of Zhengzhou University ,the medical history of 221 mothers by IVF were collected .Inclusion criteria were at least one of the parents was positive for hepatitis B surface antigen (HBsAg) ,and HBV DNA<1 × 104copies/mL . Hepatitis B vaccination status and HBV infection status of children were followed by telephone .Results One hundred and seventy‐two cases were followed up successfully ,and 49cases were lost .Two hundred and twenty‐four cases of children were HBsAg negative ,but one of them showed hepatitis B virus e antibody and hepatitis B virus core antibody positive .Conclusions The vertical transmission risk of HBV infection of children born to HBV‐positive patent(s) by IVF did not increase .

Objective To evaluate the treatment of hypervascular hepatic metastasis with TACE. Methods One hundred and twenty nine cases of hepatic metastasis treated by TACE were selected retrospectively and then analyzed the survival rate,clinical effectiveness and lipidol deposition quantity in tumor.Results Malformation of tumor vessels and rich blood supply were found in all cases of this study.The survival rates of 6 months,1 year and 3 years were 100%,73.6% and 26.4% respectively.The clinical effective rate was 68.2%(88/129)and no-progress rate was 23.3%(30/129).The satisfactory lipidol deposition quantity was obtained in 80.9%(97/129).Conclusions TACE is a favorable method for hepatic metastasis,and discerning the hypervascular subgroup could improve the treating effectiveness and be useful to make an appropriate planning.

Abstract: Objective　The method of "molecular docking - molecular dynamics - in vitro pharmacology" was used to screen and optimize small molecule polypeptide compounds that can block PD-1/PD-L1 binding and have high anti-tumor activity. Methods Using the 5-peptide compound database constructed by our team and the three-dimensional structure data of the PD-L1 protein downloaded from the protein database (PDB), the peptide compounds obtained by flexible molecular docking using Molecular Operating Environment software. The molecular dynamics calculation and analysis of the compounds with the highest GWVI/WSA (generalized Born volume integral/weighted surface area) free energy change value (ΔG) were carried out, including the root mean square deviation (RMSD) of the position change of the weight atom and the interaction energy (the sum of Lennard-Jones potential and Coulombic energy). The blocking effect of ligand compounds on PD-1/PD-L1 binding was analyzed by the homogeneous time-resolved fluorescence (HTRF) technique. The co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells was established to explore the effect of compound ligands on the killing effect of T cells on tumors and the effect on the secretion level of IL-2 in the co-culture supernatant. Results The selected polypeptide compounds were analyzed by molecular dynamics, and the binding of RGGHA to RGGHH and PD-L1 was stable. There are many kinds of interactions between RGGHA and PD-L1，which can compete with PD-1 ligands to bind Asp122, Try123 and Lys124 sites of the PD-L1 protein and block PD-1/PD-L1 signal transduction. HTRF experiments showed that the binding inhibition rate of RGGHA to PD-1 and PD-L1 was 58.38%, and that of RGGHH was 42.73%. In addition, we established a co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells to explore the immunostimulatory effect and mechanism of peptides. The results show that RGGHA and RGGHH can significantly increase the secretion level of IL-2, improve the killing ability of T cells against tumor cells and activate the tumor immune microenvironment. Conclusions The study found that the polypeptide compounds RGGHA and RGGHH can effectively block the PD-1/PD-L1 interaction and reactivate the anti-cancer immune response, which can be used as lead compounds for new drug development.

ABSTRACT:In order to explore the spatial clustering and influential factors of HFMD in Chongqing ,China from 2008 to 2012 ,spatial autocorrelation and spatial regression analysis (using the spatial lag model in this study ) were carried out using the HFMD data of 38 districts (counties) from 2008-2012 in Chongqing by OpenGeoDa ,and the HFMD case‐based data was collected from the Disease Supervision Information Management System of Chongqing Center for Disease Control and Preven‐tion .We found that the global Moran’s I coefficient of Chongqing from 2009 to 2012 was 0 .458 7 ,0 .567 5 ,0 .398 6 ,and 0 .606 0(P<0 .01) ,respectively ,indicating that the incidence of HFMD in Chongqing had a positive spatial correlation in this four years and a spatial clustering distribution was displayed .However ,no global spatial autocorrelation was founded in 2008 and the global Moran’s I coefficient was 0 .133 2 (P>0 .05) .Results of multi‐factor spatial lag regression analysis demonstra‐ted that the incidence of HFMD was positively related with urban rate (β=1 .667 6 , P=0 .001 6) ,and negatively correlated with medical technical personnel per thousand (β= -0 .000 2 ,P=0 .019 8) .In general ,the incidence of HFMD was found ge‐ographically clustered in Chongqing from 2009 to 2012 which was significantly influenced by urban rate and medical technical personnel per thousand population ,and while the urban rate was the main factor .

Objective To observe the clinical effect ofTongfu-Xiefei decoction for the patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) with phlegm-heat syndrome.Methods A total of 78 AECOPD patients were selected from June 2015 to December 2016 in the respiratory department of our hospital, and the patients were randomly divided into two groups according to the random number table method, 39 patients in each group. The control group was treated with convertional western medicine and placebo decoction. Treatment group was treated with convertional western medicine and true decoction. Both groups were treated for 2 weeks and fellowed up for 4 weeks. The symptoms score, SGRQ scores, blood gas analysis were observed and analyzed. The clinical curative effect and adverse evevts were observed.Results After treatment, the symptom cough (2.34 ± 1.11vs. 2.67±1.05,t=2.277), expectoration (1.91 ± 1.41vs. 2.35 ± 1.83, t=2.106), asthmoid (1.73 ± 1.15vs. 2.04 ± 1.53,t=2.214) scores in the treatment group were significantly lower than the control group (P<0.05). The SGRQ scores (42.47 ± 17.24vs. 59.95 ± 17.58,t=2.112) in the treatment group was significantly lower than the control group (P<0.05). The total effective rate in the treatment group was 91.9% (34/37) and 84.4% in the control group (27/32). The difference between the 2 groups was statistically significant (χ2=4.578,P=0.040).Conclusions TheTongfu-Xiefei decoction can effectively alleviate the symptoms and signs, improve the blood gas analysis index, and the long-term curative effect for AECOPD patients with phlegm-heat syndrome.

OBJECTIVETo investigate the effects of D-galactose (D-gal) on aging of rat marrow mesenchymal stem cells (MSCs) and its mechanism.

METHODSMSCs isolated from young (7 d) SD rats were randomly divided into four groups:control group, 1g/L, 10g/L and 50g/L D-gal treatment groups. In control group MSCs were cultured in DMEM containing 10% FBS for 48 h. In the D-gal treatment groups, MSCs were cultured in DMEM containing 10% FBS with 1g/L, 10g/L or 50g/L D-gal for 48 h. The senescence-associated changes were examined with SA-β-galactosidase (SA-β-gal) staining, the expressions of p53, p21 and p16 were detected by Western blot. The living and apoptotic cells were determined by AO/EB staining. Cell proliferation was detected by MTT assay. SOD activity was measured by xanthine oxidase method, and the MDA content was estimated with thiobarbituric acid (TBA) method.

RESULTSCompared to control group, the number of SA-β-gal positive cells and the expression of p53, p21 and p16 were significantly increased in the 10g/L and 50g/L D-gal treatment groups. The apoptosis rate in 50g/L D-gal group was significantly higher than that in control group (P<0.01). The proliferation of MSCs was decreased in the 10g/L and 50g/L D-gal groups compared to control group (P<0.05). After 10g/L and 50g/L D-gal treatment, SOD activity was significantly decreased (P<0.01), and MDA level was increased (P<0.01).

CONCLUSIONThe aging of MSCs can be induced by 10g/L and 50g/L D-gal, which may be associated with the elevated levels of oxidative stress.

Animals ; Apoptosis ; drug effects ; Cells, Cultured ; Cellular Senescence ; drug effects ; Galactose ; pharmacology ; Male ; Mesenchymal Stromal Cells ; drug effects ; physiology ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the safety of the da Vinci Si surgical system in thyroid surgery and to accumulate operation experience .Methods The da Vinci Si surgical system consists of a surgeon's console, a patient-side robotic cart, and high-definition 3D vision system.The robot arm tips were introduced via a single axillary incision or small chest wall ports and attached to the arms of the robot .The surgeon , sitting at the con-sole, manipulated the EndoWrist instruments and endoscope view that transfer the surgeon 's movements to the arm tips.The so called EndoWrist'technology offers seven degrees of movements ( up, down, left, right, twist, et al) , thus exceeding the capacity of a surgeon's hand in open surgery .The da Vinci Si surgical system was used to perform thyroidectomy in two small pigs .The animals were intubated and kept anesthetized with halothane .Re-sults Four surgical procedures were done using the da Vinci Si system from beginning to the end , including 2 thyroidectomies and 2 thymectomies .No conversions to open or laparoscopic procedure was used .The procedure length was 61 minutes and 50 minutes, respectively.Conclusions Robotic thyroidectomy can be performed safe-ly by the experienced surgeons after short-term da Vinci Si surgical system training .The learning curve for robotic thyroidectomy is shorter .