OBJECTIVETo observe the change of PTEN/PI3K/AKT pathway hypoxia-inducible factor (HIF-1α), vascular endothelial growth factor (VEGF) in rats with adjuvant arthritis and to explore the mechanism of neovasculization in rheumatoid arthritis.

METHODSThirty rats were randomly divided into normal control group and model control group. The model control group were established the model of adjuvant arthritis using Freund's complete adjuvant. At 19 days after modeling, the expression of microvascular density (MVD), HIF-1α, VEGF were detected by ELISA assay and PTEN, PI3K, AKT were detected by Werstern Blotting.

RESULTSCompared with the normal control group, paw swelling, arthritic index were increased, and the expression of MVD, VEGF, HIF-1α of serum, PI3K, AKT of synovial tissue were significantly increased, PTEN was significantly decreased in model control group. PI3K, HIF-1α were positively correlated with MVD; VEGF, AKT were positively correlated with paw swelling; PTEN was negatively correlated with the arthritis index; HIF-1α was positively correlated with VEGF; PI3K was positively correlated with AKT, PTEN was negatively correlated with PI3K, AKT, VEGF.

CONCLUSIONImbalance of PTEN/PI3K/AKT pathway in rats with adjuvant arthritis is one of the mechanisms of synovial neovasculization.

Animals ; Arthritis, Experimental ; physiopathology ; Hypoxia-Inducible Factor 1, alpha Subunit ; physiology ; Neovascularization, Pathologic ; etiology ; PTEN Phosphohydrolase ; physiology ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology

Objective To study the cardiotoxicity of trastuzumab in human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer patients.Methods We retrospectively collected the clinical data of general characteristic,left ventricular ejection fraction (LVEF) and clinical efficacy in 102 HER2 positive metastatic breast cancer patients treated with trastuzumab and analyzed the factors which influence the changes of LVEF and the incidence of cardiotoxicity induced by trastuzumab.Results In all of the 102 patients,the complete remission and partial remission were 7 cases (6.86％) and 73 cases (71.57％) in patients firstly treated with trastuzumab-based regimens respectively.The LVEF value was (72.85 ± 4.64) ％ at baseline and decreased to (66.05 ±5.96)％ and (65.15 ±3.38)％ as the valley points at21 and 39 months in patients who continually used trastuzumab.The progression free (PFS) of the first use of trastuzumab containing regimen and trastuzumab usage time were the clinical factors influencing the LVEF decreased value (P ＜ 0.05).The cumulative incidence of trastuzumab associated cardiotoxicity (TACT) was 14.71％ (15 cases) in the whole course of LVEF monitoring.All patients had no occurrence of heart failure and trastuzumab treatment related death caused by trastuzumab.Conclusion The heart function deceased and recovered periodically under treatment with trastuzumab in metastatic breast cancer.The incidence of cardiotoxicity was stable.Long term use of trastuzumab was well tolerated and safe.

Objective To understand healthcare-associated infection(HAI)in group burn/trauma patients,and explore its risk factors.Methods HAI occurred among 25 hospitalized patients with group burn/trauma during a bus deflagration event on May 12,2014 were monitored and analyzed.Results Of 25 burn/trauma patients,7 de-veloped 10 times of HAI,incidence and case incidence of HAI were 28.00% and 40.00% respectively;the main in-fection sites were wound and lower respiratory tract,accounting for 60.00% and 30.00% respectively.30 strains of pathogens were isolated from 7 patients with HAI,including 16 strains (53.34%)of gram-negative bacteria,13 strains(43.33%)of gram-positive bacteria,and 1 fungus (3.33% ).Risk factors for HAI were large burned area, high degree of inhalation injury,arteriovenous catheterization,urinary tract catheterization,tracheotomy,use of ventilator,and surgery.Conclusion Incidence of HAI is high in the group burn/trauma patients,corresponding in-tervention measures should be formulated according to the risk factors,so as to reduce the occurrence of HAI.

Objective To evaluate the imaging methods,characteristics,diagnostic value of multi- slice CT urography(MSCTU)in congenital abnormities of urinary system.Methods To collect 33 urinary congenital abnormities cases in three years and to analyses these MSCTU images.All examinations were performed with a multi-slice spiral CT scanner.The patients were intravenously injected with 90 ml of Iohexol 300 with a power injector at the rate of 3 ml/s.Nephrographic-phase images were obtained at 75 s after initiation of the injection of contrast material,the appropriate delay time is according to Kidney's enhancement extent and nephrohydrosis degree.Excretory-phase images were obtained through the abdomen and pelvis from 10 min.to 23 h after initiation of the injection of contrast material without abdominal compression.Excretory-phase images were transferred to the workstation and performed maximun intensity projection(MIP),multiplanar reconstruction(MPR),volume rendering(VR),and virtual cystoscopy (VC).Results The urinary congenital abnormities diagnosed by MSCTU in 33 cases,including 1 ectopic kidney,1 horseshoe kidney,1 renal malrotaion,2 supernumerary kidneys,2 ureteral valves,2 retrocaval ureters,4 congenital megaureters,6 ureteropelvic junction stenosis,9 pelviureteric duplication malformations and 5 bladder diverticula.The displaying rate of ureter was 91%(61/66).The scanning time of excretory-phase was less than 20 seconds in All cases.The average CT value of contrast media in displayed ureter lumens was 520 HU.The postprocessing images had clear,dimensional feature and It was satisfy the diagnosis.Conclusion MSCTU has clear,dimensional feature and has strong ability of displaying total anatomy shape and tiny pathology change of congenital abnormities in the urinary system.It is a very useful method for detecting the congenital abnormities in the urinary system.

Objective To investigate the CT findings of Madelung's disease in the head and neck region,and to evaluate the value of CT in demonstrating the Madelung's disease in the head and neck region.Methods CT findings of Madelung's disease in the head and neck region in 7 cases were analyzed retrospectively.All were males,with the age from 36 to 60 years,mean 51 years.All patients were underwent CT native scan,and enhanced CT scan was performed on 3 of them.Results CT images in the neck of all patients showed accumulation of nonencapsulated fat within the subcutaneous tissue and(or) deep to the platysma,and(or)within the spaces between the muscles.The fat deposits were ill-defined and symmetrical.In most cases the fat deposits involved the anterior part of the neck(infrahyoid and suprahyoid),submandibular region,the subcutaneous tissue of the nape and deep to the stenomastoid muscles.Conclusions Madelung's disease in the head and neck region have characteristic CT findings,and CT has great value in qualitative and quantitative diagnosis in Madelung's disease.

Objective To observe the influence of either alone or combined mixed-tocopherols combined with eicosapentaenoic acid (EPA) and α-Tocopherol use on oxidized LDL (oxLDL) induced 8-hydroxy-2'-deoxyguanosine (8-OHDG) and interleukin-6 (IL-6) secretion by human umbilical vein endothelial cells(HUVECs) and to explore the potential mechanism. Method Cultured HUVECs in vitro were incubated with oxLDL, oxLDL + α-tocopherol, oxLDL + mixed-tocopherols, oxLDL + EPA, oxLDL + α-tocopherol + EPA, oxLDL + mixed-tocopherols + EPA for 24 hours, respectively. Secretion of 8-OHDG and IL-6 were detected by cell enzyme linked immunosorbent assay (ELISA). The expressions of superoxide dismutase (SOD), protein kinase C-δ (PKC-δ), phosphorylated PKC-δ (p-PKC-δ) were analyzed by Western blot. Results 8-OHDG and IL-6 secretion of HUVECs was significantly increased significantly after incubated with oxLDL for 24 hours which could be significantly attenuated in the presence of tocopherols and EPA ( alone or in combination, all P ＜ 0. 05 ) while the strongest inhibition effects were seen with combined use of mixed-tocopherols and EPA. Moreover, combination of mixed-tocopherols and EPA could also significantly increase SOD activity and decrease PKC activity (all P ＜ 0. 05 ). However, the protein expression of SOD and PKC-was similar among groups. Conclusion Combined mixed-tocopherols + EPA use enhanced the inhibiting effects on the secretion of 8-OHDG and IL-6 in oxLDL stimulated HUVECs which might be linked with increased SOD activity and reduced p-PKC activity.

In this study, the inhibitory effect of human umbilical cord-derived mesenchymal stem cells (hUCMSC) on interleukin-17 (IL-17) production in peripheral blood T cells from patients with spondyloarthritis (SpA) were investigated, in order to explore the therapeutic potential of hUCMSC in the SpA. Peripheral blood mononuclear cells (PBMNC) were isolated from patients with SpA (n = 12) and healthy subjects (n = 6). PBMNC were cultured in vitro with hUCMSC or alone. The expression of IL-17 in CD4(+) T cells or γ/δ T cells were determined in each subject group by flow cytometry. IL-17 concentrations in PBMNC culture supernatants were measured by ELISA. The results indicated that the proportion of IL-17-producing CD4(+) T cells and IL-17-producing γ/δ T cells of SpA patients were 4.5 folds and 5 folds of healthy controls [CD3(+)CD4(+)IL-17(+) cells (3.42 ± 0.82)% vs (0.75 ± 0.25)%, P < 0.01; CD3(+)γδTCR(+)IL-17(+) cells (0.30 ± 0.10)% vs (0.06 ± 0.02)%, P < 0.01]. After co-culture of PBMNC in patients with hUCMSC, the increased proportions of CD3(+)CD4(+)IL-17(+) cells and CD3(+)γδTCR(+)IL-17(+) cells in SpA patients were inhibited significantly by hUCMSC [CD3(+)CD4(+)IL-17(+) cells (3.42 ± 0.82)% vs (1.81 ± 0.59)% (P < 0.01); CD3(+)γδTCR(+)IL-17(+) cells (0.30 ± 0.10)% vs (0.16 ± 0.06)% (P < 0.01]. In response to phytohemagglutinin (PHA, 1 µg/ml), PBMNC from SpA patients secreted more IL-17 than that from healthy control [(573.95 ± 171.68) pg/ml vs (115.53 ± 40.41) pg/ml (P < 0.01)]. In the presence of hUCMSC, PBMNC of SpA patients produced less amount of IL-17 [(573.95 ± 171.68) pg/ml vs (443.20 ± 147.94) pg/ml, (P < 0.01)]. It is concluded that the IL-17 production in peripheral blood T cells from SpA patients can be inhibited by hUCMSC, which have therapeutic potential for SpA.
Humans ; Interleukin-17 ; metabolism ; Leukocytes, Mononuclear ; cytology ; Lymphocyte Count ; Mesenchymal Stromal Cells ; Spondylarthritis ; blood ; metabolism ; therapy ; T-Lymphocytes ; metabolism ; Umbilical Cord ; cytology

OBJECTIVETo modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis.

METHODSSertoli cells which was isolated from 2 - 4 week-old Sprague Dawley (SD) rats, were successfully prepared by collagenase type V, trypsin and DNase I and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. The apoptosis rates of Balb/c mouse lymphocytes were examined which were co-cultured with Sertoli cells of SD rats by flow cytometry, too. The expression of FasL, TGF-beta(1) and clusterin on Sertoli cells were detected by immunocytochemistry.

RESULTSIn the co-cultured system, Sertoli cells accounted for more than 90%. The viability of Sertoli cells was above 95% and the apoptosis rate was 10.87% +/- 3.87% in this study. The lymphocytes apoptosis ratio was 15.52% +/- 0.17% (P < 0.01). Streptavidin-biotin-peroxidase-complex immunochemistry staining showed that the Sertoli cells could express FasL, TGF-beta(1) and clusterin, respectively.

CONCLUSIONSIt indicates that the expression of FasL, TGF-beta(1) on the Sertoli cells might relate to the immune privilege, and it supposed to be benefit for xenotransplantation.

Animals ; Apoptosis ; Cell Culture Techniques ; methods ; Cell Survival ; physiology ; Cells, Cultured ; Clusterin ; metabolism ; Coculture Techniques ; Fas Ligand Protein ; metabolism ; Flow Cytometry ; Immunohistochemistry ; Lymphocytes ; cytology ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron, Transmission ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; cytology ; metabolism ; ultrastructure ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo study the effect of thioridazine on the proliferation and apoptosis of human colorectal cancer SW480 cells.

METHODSSW480 cells were treated with different concentrations of thioridazine, and MTT assay was used to evaluate the cell inhibition rate. Hoechst 33342 staining was performed to demonstrate the cell morphology changes. Flow cytometry was used to determine the cell apoptosis and cell cycle changes. RT-qPCR was used to detect PDCD4, c-MYC, BCL2, CCND1, CASPASE3, PARP1, CDK4 and EIF4A mRNA expressions, and Western blotting was employed to assay AKT, p-AKT, and PDCD4 protein expression levels.

RESULTSMTT results showed that thioridazine inhibits the proliferation of SW480 cells. SW480 cells treated with thioridazine presented with such typical features of apoptosis of karyopyknosis, chromatin condensation and nuclear fragmentation. Flow cytometry showed that thioridazine was a cell cycle-specific drug and caused cell cycle arrest at G(1)/G(0) phase and an increased cell apoptosis rate. Thioridazine treatment of the cells resulted in up-regulated PDCD4 mRNA expression and down-regulated mRNA expressions of CCND1, CDK4, c-MYC, BCL2, CASPASE3, PARP1 and EIF4A, increased PDCD4 protein expression and reduced p-AKT protein expression.

CONCLUSIONThioridazine inhibits the proliferation and induces apoptosis of SW480 cells by up-regulating PDCD4 and inhibiting PI3K/Akt pathway.

Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Cycle Checkpoints ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Down-Regulation ; Humans ; RNA-Binding Proteins ; metabolism ; Signal Transduction ; drug effects ; Thioridazine ; pharmacology

OBJECTIVETo evaluate contrast-enhanced ultrasonography (CEUS) in detecting testicular perfusion in acute testis contusion.

METHODSWe established the model of testis contusion in 11 healthy male New Zealand rabbits by randomly hitting one side of the scrotum under general anesthesia. We examined the bilateral scrotums of all the animals before, immediately after and at 2, 4 and 6 hours after modeling by color Doppler flow imaging (CDFI) and CEUS, and analyzed the time-intensity curve (TIC), arriving time (AT), time to peak intensity (TTP), peak intensity (PI), half time of descending peak intensity (HT) and area under the curve (AUC) in the healthy and injured testis, respectively.

RESULTSCEUS exhibited a higher sensitivity in detecting tissue perfusion than CDFI. The mode of contrast agent perfusion in testicular contusion was fast in and slow out. There were no evident differences between the contused and the healthy testis in AT, TTP and PI before modeling. The contused testis showed significantly earlier AT and TTP, higher PI and larger AUC (P < 0.05) than the healthy one at different time points after modeling, but no statistically significant difference was found in HT (P > 0.05).

CONCLUSIONAccurate parameters of testicular perfusion in acute testis contusion can be quantitatively obtained by CEUS, which are of important value for the diagnosis of testis contusion.

Animals ; Contrast Media ; Contusions ; diagnostic imaging ; Male ; Rabbits ; Testis ; blood supply ; diagnostic imaging ; injuries ; Ultrasonography, Doppler, Color