Objective To determine the modality of Leptospira interrogans invading human and murine mononuclear-macrophages and diversity of leptospiral phagocytotic vesicle formation. Methods Transmission electron microscopy was applied to observe the invasion of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai into murine mononuclear-macrophage-like cell line J774A. 1 and PMA-activated human monocyte line THP-1 and the formation of leptospiral phagocytotic vesicles. By using immunofluorescence plus either laser confocal microscopy or fluorescence spectrophotometry, the changes of intracellular leptospiral numbers in J774A. 1 and PMA-activated THP-1 cells before and after block with endocytosis inhibitors monodansylcadaverin (MDC), phenylarsine oxide (PAO) and clathrin antibody were investigated. Results The leptospires in J774A. 1 cells were located in phagocytotic vesicles while the leptospires in THP-1 cells had no package with phagocytotic vesicle membrane. Both MDC and PAO presented the effect inhibiting endocytosis of L. interrogans into J774A. 1 and THP-1 cells in dose-dependent manner. The numbers of leptospires in J774A. 1 and THP-1 cells that pre-blocked with 10 μmol/L or above MDC and 1 μmol/L or above PAO were significantly less than that in the two cells untreated with MDC and PAO (P＜0.05＝. After J774A. 1 and THP-1 cells were blocked with clathrin antibody, the numbers of intracellular leptospires were also remarkbly decreased ( P<0.05 ).Conclusion Leptospira interrogans can invade into both human and murine mononuclear-macrophages through the way of clathrin-dependent endocytosis. There is an opposite diversity of leptospiral phagocytotic vesicle formations in human and murine mononuclear-macrophages, which may result in the difference of pathogenesis in human and mice after infected with L. interrogans.

BACKGROUND:Pathological mechanism of femoral head necrosis is not fuly known, and high-dose corticosteroids are prone to induce femoral head necrosis.
OBJECTIVE:To investigate the clinical effect of bone marrow mesenchymal stem cels combined with theaflavin on steroid-induced avascular necrosis of femoral head in rats.
METHODS: Rat bone marrow mesenchymal stem cels were isolated and culturedin vitro and compounded onto gelatin sponge, and then, divided into four groups: model group, decompression group, cel transplantation group and combined group. Models of steroid-induced avascular necrosis of femoral head were established using liquid nitrogen method. In the combined group, gelatin sponge carrying bone marrow mesenchymal stem cels was transplanted and 250 mg theaflavin was given daily after modeling.
RESULTS AND CONCLUSION:At 4 weeks after modeling, the femoral head samples in the four groups were round with exfoliation of articular cartilage that was pale in color. Then, the exfoliation of articular cartilage was aggravated in the model group, and some specimens appeared to colapse; the femoral head specimens in the combined group were round and pale. Until the 8th week after modeling, bone necrosis was increased in the model group; osteoblasts were found in the decompression group with fibrous calus formation; a few of empty bone lacunae appeared with irregular medulary cavity in the cel transplantation group; a large amount of new bone tissues formed in the combined group, and intramedulary adiocytes were in regular array. At 4 and 8 weeks after modeling, the number of empty bone lacunae was significantly lower in the combined group than the other groups (P < 0.05). These findings indicate that bone marrow mesenchymal stem cels combined with theaflavin is an ideal therapy for steroid-induced avascular necrosis of femoral head.

Objective To explore the effect of ?-elemece on the reverse of drug resistance and the expression of P-glycoprotein(P-gp) in MDR K562/ADM cells. Methods MTT method was used to detect the sensitivity of cell and its reverse of drug resistance.Immunohistochemistry and immunoelectron microscopy were for observing the expression of P-gp.It's quantitative analysis was determined by flow cytometry.All experimental data were dealt with by SPSS(10.0 production pacility) soft ware. Results 1.Non-cytotoxic dose of ?-elemece(4.0?mg/L) could obviously decrease the IC-(50) value of K562/ADM cells to ADM.The reversing fold was 2.18;2.The level of P-gp expression was higher in K562/ADM cells than that in its parent K562 cells.Our studies also showed that the distribution of P-gp was on membrane of K562/ADM cells;3.Non-cytotoxic dose of ?-elemece(4.0?mg/L) could remarkably decrease the P-gp expression in MDR K562/ADM cells.Conclusion\ ?-elemece could evidently reverse drug resistance of K562/ADM to ADM.Decreasing the P-gp expression in drug resistance cell is one of the main mechanism of reversion.

Objective To explore the role of EphB4 in proliferation of glioma cells. Methods The mRNA and protein expressions of EphB4 were detected using RT-PCR, immunochemistry, and Western-blot, respectively. EphB4 siRNA was synthesized and transfected into U251 cells using Lipofectamine 2000. Glioma cell proliferation, apoptosis, and invasion were determined by MTT assay, TUNEL and transwell experiment, respectively. Results The expression (P<0.05) and proliferation of EphB4 were obviously decreased in U251 transfected with EphB4 siRNA and the proliferation was further decreased with the increased concetrations of siRNA. Compared with U251 group and siRNASCR group, EphB4 siRNA at different concentrations (25, 50 or 100 nmol/L) significantly reduced the invasion ability of cells and increased the number of apoptotic cells (P<0.05). Conclusions EphB4 plays an important role in the regulation of glioma cell proliferation, apoptosis and invasion.

Abstract A new method was established to determine the free inorganic cadmium ion ( Cd2+) in marine bivalves using high performance liquid chromatography and inductively coupled plasma mass spectrometry ( HPLC-ICP-MS) . The free Cd2+ in shellfish was ultrasonic extracted for 40 min by 10 mmol/L Tris buffer solution ( with 0. 1 mol/L NaCl, pH=7. 5), and the separation of Cd2+ was achieved using an IonPac CS5A analytical column with an IonPac CG5A guard column. The mobile phase consisted of 50 mmol/L C2 H2 O4 and 95 mmol/L LiOH. The new method had a good linear relationship with the correlation coefficient of 0. 999 and the standard recoveries of Cd2+ were all above 84. 6%. The free Cd2+ content in several kinds of marine bivalves was determined using the method and the results showed that the percentage of free inorganic Cd2+ to the total Cd content in samples with high Cd content was higher than those with low Cd content.

Objective:To ascertain whether the immune complexes (ICs) formed by Dengue virus 1 non-structure protein 1 (DENV1 NS1)and its IgG antibodies could mediate passive systemic anaphylaxis (PSA) and to explain the pathogenesis of Dengue hemorrhagic fever or Dengue shock syndrome (DHF/DSS).Methods:The monoclonal antibodies (mAbs) or mAb cocktails from 20 IgG mAbs of DENV1 NS1 prepared in this lab were screened to initiate PSA and passive cutaneous anaphylaxis (PCA) in mice.Meanwhile, the effects of GdCl3 and platelet activating factor ( PAF) antagonist CV-3988 on PSA induced by the NS1-IgG ICs were observed.Results:Two groups of monoclonal antibody cocktails with purified NS 1 were proved to be capable of provoking PCA and PSA in mice,whereas the other mAbs or mAb cocktails could be not .The murine PSA initiated by NS1-IgG(5D25+3B1) ICs could be sig-nificantly inhibited by in vivo treatment with GdCl3 or PAF antagonist CV-3988.Conclusion: The NS1-IgG ICs formed with DENV1 NS1 and IgG mAb cocktails can mediate PSA and PCA ,but not all of ICs formed by DENV 1 NS1 mAbs or mAb cocktails with DENV 1 NS1 can induce PSA ,indicating that it may be related to the special epitopes of DENV 1 NS1.The monocyte/macrophages and PAF may be as major effector cells and the major mediator for PSA induced by NS 1-IgG ICs,respectively.

0.05). The generation of ROS increased obviously after the cells were incubated with them for 12 h, and the increase of ROS reached the peak treated for 24 h. The levels of ?? m were time-dependently decreased after treating with four compounds for 12, 24, and 48 h. Conclusion The grouth of both MDR KBv200 cells and parental drug-sensitive KB cells were inhibited to the treatment of four anthraquinone derivative in vitro. The mechanism of their effects is associated with the increase of the cellular ROS level and the decrease of ?? m.

Background The advantages of endothelium keratoplasty(EK)include without corneal surface incision and sutures,remaining normal corneal topography,minimizing astigmatism and providing a healthy donor endothelial cell count and function.Using anterior segment optical coherence tomography(AS-OCT) to examine the morphology of the corneal graft after EK is helpful for the evaluation of the outcome of EK.ObjectiveThis study applies AS-OCT to observe the relationship of recipient cornea and donor graft and offer a basis for the evaluation of operative outcome.MethodsThe clinical data from 44 eyes of 42 patient who received DSAEK in Peking University Eye Center from September 2007 through April 2009 was reviewed retrospectively.These patients included 30 male and 12 female with the age from 15 to 83 years.Structure of cornea and relationship between recipient cornea and graft disc after surgery were clinically examined and recorded by slit-lamp digital camera and AS-OCT.This study was approved by the Ethics Committee of Peking University.Written informed consent was obtained from all of the patients before the operation.ResultsThe donor graft attached to posterior central surface of recipient in 28 eyes under the slit-lamp microscopy and AS-OCT.The thickness of donor graft was decreased,and graft sticked more closely to recipient cornea and was more transparent with the lapse of time.Complete dislocation of the donor graft in early stage was found in 5 eyes(11.36%) and partial dislocation in 2 eyes(4.55%).Curliness of graft edge occurred in 1 eye(2.27%) and eccentricity of donor graft in 8 eyes(18.18%).ConclusionAS-OCT can display the morphology and structure of cornea as well as the relationship between donor graft and recipient after DSAEK.It is helpful for evaluating the clinical effectiveness of DSAEK.

Objective:To study the effectiveness and safety of combined intraoral and cutaneous using of long pulsed 1064 nm Nd∶YAG and 2940 nm Er∶YAG lasers in facial rejuvenation.Methods:Twenty-one female patients with different grades of nasolabial fold were treated with 6 sessions of the combination lasers therapy once a month. Standardized photographs of VISIA were taken before and after treatment to record any changes of facial contour and nasolabial folds. The efficacy assessment was judged by two non-group experienced dermatologists using modified Fitzpatrick Wrinkle Scale (MFWS), based on the VISIA photos at base line, twelve-week and six-month after treatment. Self-scoring of 5-level satisfaction from patients was also collected and analyzed. Any side effects and complications were recorded.Results:At 12 weeks, mild and moderate nasolabial folds were improved notably, the mean value of MFWS decreased from 1.59 to 0.90, whereas severe nasolabial folds had no remarkable improvement. Six-month after treatment, 18 patients had notable improved, among them: five out of 7 mild cases, 9 out of 10 moderate cases and 2 out of 4 severe cases, and the mean value of MFWS significantly decreased to 0.57 ( P<0.001). The general satisfactory rate of self-scoring was 86%. Two patients had tolerable light burning pain during treatment and two patients got temporary pigmentation that vanished in three months. There were no other adverse reactions and complications. Conclusion:Combined intraoral and cutaneous using oflong pulse 1064 nm Nd∶YAG laser and 2940 nm Er∶YAG laser is a safe and effective therapy for facial rejuvenation.