Objective Previous researches demonstrated that pyrroledithiocarbomate (PDTC), an inhibitor of nuclear factor, results in specific inhibition on nuclear factor-κB and therefore suppress cataract formation. The aim of this study was to explore the effect of nuclear factor-κB inhibitor on cataract formation after alkali burn in rats. Methods Ocular surface alkali burn models were established in the right eyes of 40 SPF Wistar rats by putting the 7 mm filter paper with 1 mol/L NaOH in the central cornea for 60 seconds. PDTC (2 mg/mL, 0. 1 mL) was subconjunctivaly injected everyday in 20 model eyes and the equivalent amount of normal saline solution was used in the same way in other 20 model eyes. The rats were killed in the first, third, fifth and seventh day after alkali burn and lenses were obtained for the histopathological examination, and immunochemistry and polymerase chain reaction were used to detect the expression of nuclear factor-κB in lens epithelial cells. The experiment and use of animal followed the Standard of Association for Research in Vision and Ophthalmology. Results After alkali burn, lens epithelial cells namely fell off in control group but those of experimental group were complete in the first day. In the third day, the lens cortex was obviously condensed in both two groups. In the fifth and seventh day, the lens epithelial cells fell off and lens cortex was obviously briquetted. Lots of vacuole and fragments could be seen in both groups. The gray scale value of nuclear factor-κB in lens epithelial cells were significantly higher in 1 day and 3 days after alkali burn in PDTC group compared with control group(t =2. 836, P =0. 036; t =4. 932, P =0.004) . The nuclear factor-κB /p-actin values were considerably lowed in 1 day and 3 days after alkali burn in PDTC group compared with control group (t = 31. 563, P = 0. 000; t = 17. 837, P = 0. 000). No statistically significant difference were found in gray scale values of nuclear factor-κB and nuclear factor-κB/p-actin values in 5 days and 7 days after alkali burn between PDTC group and control group (P＞0. 05). Conclusion Early usage of inhibitor of nuclear factor plays a suppressive role in cataract formation after ocular surface alkali burn of rat model.

Objective:To explore the methods and condition in which calcium ionorphore(CI) induces the CML cells to differentiate into dendritic cells(DCs).Methods:Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30?30~9 L~ -1 when samples were collected,then lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10%FCS(Fetal calf serum),with or without CI(375 ng/ml) and GM-CSF(200 ng/ml) at 37℃,5%CO_2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI.Results:After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80,CD86,CD40,CD86 and HLA-DR.Conclusion:CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.

AIM and METHOD: The relationship between the evolution of the reticulin fibres(RF) or the collagen fibres(CF) and the growing of hematopoietic cells in long-term bone marrow culture (LTBMC) from 15 paticnts with acute myeloid leukemia (AML) and form 6 normal subjects was observed by inverted microscope, Gomoris staining and Massons staining were used. RESULTS: (1)The amount of RF contents of 8 AMLs,with self- maintained(AMLsm) in the 1～8 weeks-old-culture was significant less than that of normal control and 7 AMLs,without self-maintained(AMLnsm) ( P

Objective To understand the incidence of hemolytic disease in newborn (HDN)among the newborns with jaundice and the coincidence degree of the blood group serological results and the clinical diagnosis in HDN.Methods The microcolumn gel method was adopted to detect the 3 serological indexes in 276 jaundice newborns of maternal fetal blood group incompatibility,in-cluding the direct antiglobulin test,free antibody test and antibody release test.Results 108 cases of HDN were clinically diagnosed with the positive rate of 39.13%.The positive detection rate in newborns with 0-2 d old was highest(50.00%).Conclusion The serological test can provide the basis for the early diagnosis and treatment of HDN.Collecting the specimen as early as possible can improve the positive diagnosis rate of HDN.

Objective To investigate the cytokine spectrum and henlatopoiesis-supportive function of umbilical cord derived mesdnchymal stem cells(UC-MSC),and compare with those of normal adult bone marrow derived mesenchymal stem cells(BM-MSC).Methods The mRNA of cytokine production of UC-MSC and BM-MSC were determined by reverse transcriptasc polymerase chain reaction(RT-PCR)analysis.To evaluate hematopoiesis supporting activity,cord blood(CB)CD+34 cells were co-cultured with UC-MSC or BM-MSC.Colony-forming cells(CFC)were determined after 5 weeks of culture.Results RT-PCR assay showed that UC-MSC had a cytokine spectrum very similar to that of BM-MSC.including expression of the mRNA ofstem cell factor,leukemia inhibitor factor,macrophage colony stimulating factor,Flt3-ligand,interleukin-6,vascular endothelial growth factor and stromal derived factor-1.but UC-MCS additionally expressed mRNA of granulocyte macrophage and granulocyte colony-stimulating factors.After co-culture with CD+34 cord blood cells for 5 weeks,no significant difference in CFC was observed between the CD+34 cells/UC-MSC and CD+34 cells/BM-MSC co-cultures (P＞0.05). Conclusion The cytokine spectrum and hematopoiesis-supponive function of UC-MSC ale similar with that of BM-MSC.

Objective:To investigate the effects of induced pluripotent stem cell-derived exosome (iPSC-Exo) on releasing inflammatory factors from microglia induced by lipopolysaccharide (LPS).Methods:iPSC derived from the tubular epithelial cells of sepsis encephalopathy patients were resuscitated and cultured. The iPSC-Exo was isolated by low-temperature ultracentrifugation and analyzed by transmission electron microscopy, Western blot and high sensitivity flow cytometry (HSFCM). Based on the concentration of iPSC-Exo, human microglia line HMO6 cells activated by LPS (100 ng/mL) were divided into four groups randomly: LPS+ phosphate buffer solution (PBS) group, LPS+iPSC-Exo 10 5 group, LPS+iPSC-Exo 10 6 group and LPS+iPSC-Exo 10 7 group. The control group was added equal PBS but not LPS. After culture for 24 h, the concentrations of malondialdehyde in cells were detected. Quantitative RT-PCR was used to measure the mRNA expression levels of macrophage inflammatory protein 2 (MIP2), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 in the cells and enzyme-linked immunosorbent assay (ELISA) was used to assess the concentration of these cytokines in the supernatant. Under the same concentration of iPSC-Exo, one-way ANOVA and SNK- q test were used for comparison between groups. Results:The extracts showed spherical membrane structure by transmission electron microscopy. HSFCM showed the mean diameter of the extracts was (74.66±15.60) nm and the concentration around 2.98×10 10/mL. Western blot analysis showed high expression of exosome markers CD63, Alix and TSG101, but not GM130. Intracellular MDA concentration and mRNA expression levels and protein concentration of MIP2, TNF-α, IL-1β and IL-6 in the LPS+PBS group were significantly higher than those in the control group (all P<0.01). With the increase of iPSC-Exo concentration, the intracellular MDA concentration decreased gradually ( P<0.01), the mRNA expression levels of inflammatory factors showed a gradual downward trend (all P<0.05). Each inflammatory cytokine in the supernatant declined in a manner that was almost consistent with mRNA. Concentrations of MDA remained constant in the control group. Conclusions:iPSC-Exo derived from the tubular epithelial cells of sepsis encephalopathy patients alleviate oxidative stress and inflammation effect of microglia induced by LPS, and the modulatory effect is dose-dependent.