50%).RT-PCR results were classified as positive and negative according to the presence or absence of corresponding electrophoretic bands. Results Immunohistochemically,integrin ?2 and ?3 subunits were all expressed strongly in 12 cases of normal bladder tissues (100%).In 53 cases of BTCC tissues,?2 was strongly positive in 10 cases (19%),positive in 22(42%),negative in 21(40%);?3 was strongly positive in 16 (30%),positive in 24 (45%),negative in 13(25%).The expressions of ?2 and ?3 were significantly different between normal and BTCC tissues ( P 0.05).RT-PCR results showed that ?3 expression was positive in all normal tissues (100%) and in 36 (68%) out of the 53 cases of BTCC tissues;the difference was significant ( P 0.05),but ?3 mRNA positive expression was significantly related to clinical stages [T is ～T 1:83%(19/23),T 2～T 4:57%(17/30); P

Objective:To investigate the mechanism of sinomenine (SIN) in inducing the immunosuppressive effects of rat-derived dendritic cells (DCs).Methods:The bone marrow-derived precursor cells from Wistar rats were cultured in vitro. The morphological differences between sinomenine treated DCs (sinomenine modified group, SIN group) and conventional induced DCs (conventional induced group, control group) were observed under microscope. The CD phenotype of DCs was detected by flow cytometry. DCs were induced maturation by lipopolysaccharides (LPS) stimulation. The impact of SIN on the expressions of Toll-like receptor (TLR)2, TLR3 and TLR4 on the DCs surface were detected by flow cytometry. Results:In the conventional induction group, the cells showed clusters or suspension growth, with obvious granular sense on the cell surface; while in the sinomenine induction group, the cells were clustered together, with no significant change in cell volume and morphology. The relative expressions of CD80 and CD86 were 70.7% and 71.3% in the conventional induction group, while 51.7% and 49.4% in the SIN group. The relative expression of TLRs on DCs in SIN + LPS group was TLR2 (51.2±0.34)%, TLR3 (50.3±0.14)%, TLR4 (52.1±0.16)%, which were significantly lower than those in LPS group [(94.35±0.16)%, (97.55±0.16)%, (94.6±0.12)%].Conclusions:SIN may induce immune tolerance by inhibiting the maturation of DCs via inhibiting the TLRs signaling pathways.

In this study we performed Tissue Doppler Imaging (TDI), two-dimensional speckle tracking imaging (2D- STI) and three-dimensional speckle tracking imaging (3D-STI) on enrolled healthy, overweight and obese groups (34 subjects in each group), respectively, to analyze cardiac structure and its function. Compared with healthy group, global longitudinal strain (GLS), global circumferential strain (GCS), global area strain(GAS) and global radial strain (GRS) decreased progressively (P < 0.05). The ratio of early diastolic mitral inflow velocity to global early diastolic strain rate of left ventricle (E/e'sr) (r = 0.466, P < 0.001), GLS (r = 0.502, P < 0. 001), GCS (r = 0.426, P < 0.001), GAS (r = 0.535, P < 0.001) and GRS (r = -0.554, P < 0.001) were correlated with body mass index (BMI). E/e'sr (r = 0.37, P = 0.003), GLS (r = 0.455, P < 0.001), GCS (r = 0.282, P = 0.02), GAS (r = 0.412, P < 0.001) and GRS (r = -0.471, P < 0.001) were correlated with free fatty acid (FFA). Stepwise multiple linear regression revealed that BMI was independently correlated with E/e'sr, GLS, GCS, GAS and GRS. Waist to hip ratio (WHR) was independently correlated with GLS, GCS, GAS and GRS. FFA was independently correlated with E/e'sr (P < 0.05). The study showed that cardiac structure changed and impaired left ventricular global systolic and diastolic function in overweight and obes population. Moreover, BMI, WHR and FFA may be independent influence factors of cardiac function in overweight and obese population.
Body Mass Index ; Case-Control Studies ; Echocardiography, Three-Dimensional ; Heart ; physiopathology ; Heart Ventricles ; physiopathology ; Humans ; Linear Models ; Obesity ; physiopathology ; Overweight ; physiopathology ; Reproducibility of Results ; Ventricular Function, Left

Objective To analyze the expression feature and function of microRNAs in exosomes secreted by leukemia cells (LCEX). Methods The mice leukemia cell line L1210 was taken as the example, and LCEXL1210 was obtained by isolating supernate of L1210 cells through density gradient centrifugation. MicroRNAs isolated from LCEXL1210 were analyzed by microarray analysis, compared with miRNA from L1210 cell line, and then some of miRNAs with different expression were verified by real-time PCR and were analyzed by Gene Ontology (GO) database. Results The number of miRNAs identified in LCEXL1210 was 1 044, and that in L1210 cell line was 872. The number of shared miRNAs between LCEXL1210 and L1210 cell line was 732, accounting for 70.1 % of LCEXL1210 and 83.9 % of L1210 cell line, respectively, which indicated that 70 % of LCEXL1210 was derived from the parental cells. Interestingly, 312 miRNAs in LCEXL1210 were found to be underrepresented in the parental cells, indicating their specificity in LCEXL1210. Some miRNAs were significantly highly expressed in LCEXL1210 compared with those in L1210 cell line, including miR-16-1, miR-210, miR-195 and so on, which showed that miRNAs isolated from LCEXL1210 were differentially expressed with those from the parental cells. Some differentially expressed miRNAs from LCEXL1210 were verified by real-time PCR, and then were analyzed by GO database, which demonstrated that these highly expressed miRNAs participated in the processes of various biological function and signal transduction. Conclusions MiRNAs isolated from LCEXL1210 show a high similarity to miRNAs isolated from L1210 cells, whereas of which one-third are specific. The highly expressed miRNAs participate in the processes of various biological function and signal transduction.

BACKGROUND:It may be an important approach to avoiding organ transplant rejection by utilizing immature dendritic cells to induce donor-specific immunologic tolerance. OBJECTIVE:To study the effect of sinomenine on the differentiation and maturation of rat bone marrow-derived dendritic celsin vitro. METHODS:Bone marrow-derived dendritic cells were isolated from the rat femur and tibia, and immature dendritic cells were induced by granulocyte-macrophage colony stimulating factor and interleukin-4. On day 7, lipopolysaccharide was added and the cells were cultured to generate mature dendritic cells. Cells were divided into control group and low-, middle- and high-dose sinomenine treatment groups (SNL, SNM, SNH groups). Forty hours later, dendritic cels were harvested, and cell morphology was observed by inverted phase contrast microscope. The expression of CD80 and RT1B was detected by flow cytometry. ELISA was used to detect the expression of interleukin-12. The mixed lymphocyte reaction was used to detect the ability of dendritic cells to stimulate the activation of allogeneic T lymphocytes. RESULTS AND CONCLUSION: (1) Under the inverted microscope, the morphology of mature dendritic cells was observed in the control group; in the SNL group most dendritic cells were visible; in the SNM group, there were partially suspended cells with poor maturation; and in the SNH group, most of the cells were not mature. (2) The expression of CD80 in the control group was significantly lower than that in the SNL, SNM and SNH groups (P < 0.05), and the expression of RT1B was significantly reduced in the SNM and SNH groups than the control group. (3) Compared with the control group, the level of IL-12p70 in the cell supernatant was significantly decreased in the SNM and SNH groups (P < 0.01). (4) The ability of dendritic cells to stimulate T lymphocyte proliferation in the SNM and SNH groups was significantly decreased compared with the control group (P < 0.05). To conclude, sinomenine can inhibit the maturation of dendritic cells.

Objective: To explore the mechanism of traditional Chinese medicine(TCM) combined with regular therapy in the treatment of pneumonia with systemic inflammatory response syndrome(SIRS) in(children).Methods: Sixty-four children with pneumonia including 51 cases with SIRS complication were(randomly) divided into three groups: treatment group(regular therapy+TCM,n=26),control group Ⅰ((regular)(therapy) only,n=25) and the control groups Ⅱ(13 pneumonia children without SIRS(receiving)(routine) treatment). The dynamic changes of tumor necrosis factor-?(TNF-?),interleukin-6(IL-6) and IL-8 were measured before and after treatment in all children.Serum Creactive protein(CRP) and total white blood cell(WBC) count were measured simultaneously.Clinical symptom and sign scores were(performed)(5 days) after treatment in children with pneumonia and SIRS complication.Results: Before(treatment),serum TNF-?,IL-6,IL-8 and CRP concentrations were significantly higher in treatment group and(control) group Ⅰ than(control) group Ⅱ(all P

Objective To investigate the effect of immature dendritic cell (imDC) derived from donor-derived bone marrow induced by alkaloid sinomenine (SN) on the Th1/Th2 cytokines in venous blood of receptors,and to probe into the mechanism which imDC induced by SN can lead transplantation immune tolerance.Methods Inbred strain Wistar and Sprague Dawley (SD) rats were selected as kidney transplant donor and recipient,respectively.Vessel sutures of the microsurgery technique were used to build the bilaterally renal transplantation model of rats.By the injection of imDC to the recipient rats preoperatively,enzyme-linked immunosorbent assay (ELISA) was used to determine the level of the interleukin (IL)-2,IL-4,IL-10 and interferon-γ (INF-γ).Results (1) Successful rate of transplantation was 89.5％.Arterial anastomosis time was (12.5 ±5.7)min,and venous anastomosis time was (17.3 ± 3.4)min.(2) The content of the IL-2,INF-γ,IL-4 and IL-10 in SN-imDC 106 group was (17.25 ± 3.41) pg/ml,(239.80 ± 9.06) pg/ml,(337.60 ± 25.07) pg/ml,and (1 432.00 ± 106.39) pg/ml,respectively.Among the same concentration,the level of the IL-4 and IL-10 that stood for the Th2 cytokines was significantly higher in SN-imDC group than imDC group and control group (P ＜ 0.05),and was significantly higher in SN-imDC 106 group than SN-imDC 105 group (P ＜ 0.05).Among the same concentration,the lever of IL-2 and INF-γ that stood for the Th1 cytokines was significantly lower in SN-imDC group than imDC group and control group (P ＜ 0.05),and was significantly lower in SN-imDC 106 group than SN-imDC 105 group (P ＜ 0.05).Conclusions (1) The use of microsurgery for anastomosis could make the model of singel kidney transplantation in rats.(2) Specific imDC induced by SN could induce the migration to Th2 immune response,which proved imDC induced by SN could mediate immune tolerance to the recipient.

We sought to explore the feasibility of global area strain to assess left ventricular global systolic function in patients with essential hypertension and normal ventricular geometry. Thirty-five essential hypertensive patients with normal ventricular geometry and 30 normally healthy persons as controls were enrolled in this study. The two groups were comparable for age, sex ratio, height, weight, body mass index (BMI), and heart rate. Blood pressures (BPs) were significantly higher in the hypertension group than the control group. Two-dimensional echocardiography and three-dimensional speckle tracking imaging were performed. Left ventricular global area strain (GAS), global longitudinal strain (GLS), global circumferential strain (GCS), global radial strain (GRS), LV volumes, ejection fraction (EF), sphericity index (SPI), left ventricular end-diastolic and end-systolic mass (EDmass and ESmass) and LV mass index (EDmassI and ESmassI) were obtained. Compared with those of the controls, GAS, GLS, GCS, GRS were significantly reduced in hypertensive patients Call P < 0.001). GAS (r = 0.672, P < 0. 001), GLS (r = 0.587, P < 0.001), GCS (r = 0.639, P < 0.001) and GRS (r = 0.685, P < 0.001) were correlated with EF in the pooled population. GAS showed an excellent correlation with GCS (r = 0.905, P < 0.001), GLS (r = 0.892, P < 0.001) and GRS (r = 0.990, P < 0.001). EF measured with 3D-STI was significantly lower in group of hypertension (P < 0.001) than that in the controls. There were no significant differences between the two groups in cardiac output, sphericity index, EDmass and ESmass, LV mass index (EDmassI and ESmassI) calculated with 3D-STI. The study showed that GAS could identify early changes of left ventricular global systolic function in hypertensive patients with normal ventricular geometry.
Adult ; Aged ; Case-Control Studies ; Echocardiography, Three-Dimensional ; methods ; Female ; Humans ; Hypertension ; diagnostic imaging ; physiopathology ; Male ; Middle Aged ; Myocardial Contraction ; physiology ; Ventricular Function, Left ; physiology ; Young Adult

Objective To compare the influences of microsurgical clipping and intravascular interventional embolization on the nerve function recovery in patients with posterior communicating aneurysm combined with oculomotor paralyses.Methods Eighty-six patients with posterior communicating artery aneurysm accompanied by oculomotor nerve palsy admitted to Qianjiang Central Hospital of Chongqing from May 2015 to May 2017 were randomly divided into observation group (n =43) and control group (n =43) by random number table method.The control group was treated with microsurgical clipping,while the observation group was treated with intravascular interventional embolization.The therapeutic effect,neurological function recovery,postoperative complications and prognosis of the two groups were compared.Results The operation time,intraoperative blood loss,postoperative respiratory recovery time and postoperative extubation time of the observation group were significantly less than those of the control group (P ＜ 0.05);the complete recovery of the observation group was significantly higher than that of the control group,and no recovery was significantly lower than that of the control group (P ＜ 0.05);the incidence of complications in the observation group was significantly lower than that in the control group (P ＜ 0.05);the proportion of 5 points in the observation group was significantly higher than that in the control group (P ＜ 0.05).Conclusions Intravascular interventional embolization in the treatment of posterior communicating aneurysm combined with oculomotor paralyses can promote the recovery of nerve function,reduce the incidence of postoperative complications,have a good prognosis and a definite effect,which can be used as the first choice of treatment.

Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.