Objective To explore the effects of Interleukin 10 (IL 10) on the endothelial damage mediated by inflammatory cytokines in Kawasaki disease (KD).Methods Human umbilical vein endothelial cells were cultured in vitro and the production of tumor necrosis factor alpha (TNF ?),interleukin 1 beta (IL 1?),interleukin 6 (Il 6) and IL 10 were measured by enzyme linked immunosorbent assay (ELISA).The proportion of apoptotic cells in endothelial cells induced by the supernatants of peripheral blood mononuclear cells (PBMCs) was detected by Annexin V/PI double staining though flow cytometry.Electrophoretic mobility shift assay (EMSA) was used to detect the activity of nuclear factor ?B (NF ?B).Results The level of TNF ?,IL 1?,IL 6 and IL 10 were markedly increased in KD patients and the proportion of apoptotic cells in endothelial cells induced by the cultured supernatants of PBMCs was markedly elevated (38 4?7 8)% compared to (2 8?0 8)% in the control subjects.The apoptosis of endothelial cells induced by the cultured supernatants of PBMCs could be reversed to some degree by anti TNF ?,IL 1? and IL 6 monoclonal antibody (McAb),otherwise could be enhanced (58 6?14 6)% by anti IL 10 McAb.The activity of NF ?B in the activated PBMCs in KD patients was distinctly increased and IL 10 could significantly inhibit the activity of NF ?B,reduce the production of TNF ?,IL 1? and IL 6 and the apoptosis of endothelial cells induced by the cultured supenatants of PBMCs in KD patients.Conclusion IL 10,which can inhibit the activity of NF ?B and the transcription of inflammatory cytokines such as TNF ?,IL 1? and IL 6,acts as an important protective factor in endothelial damage of KD.

AIM: To investigate whether efficient production of neuron-like cells from embryonic stem cells(ESCs) can be indued by astrocyte-conditioned medium(ACM) in vitro.METHODS: Based on the 4-/4+ protocol established by Bain,two groups were studied: ATRA group,ATRA with ACM group.ESCs were induced into neuron-like cells by means of three-step differentiation in vitro.The totipotency of ESCs was identified by the observation of cells' morphology and the formations of teratoma in immunocomprised mice.The cell differentiation was evaluated continuously by detection of the cellular specific markers of neural stem cell, neurons and astrocytes such as nestin,NF-200,NSE and GFAP using immuno-histochemistry assay.RESULTS:(1) The ESC-D3 cells kept the ability of differentiation into cellular derivations of all three primary germ layers after continuous passage culture.(2) The ratio of NF-200 and NSE positive cells in the cells induced by ATRA with ACM was higher than that in the cells induced by ATRA only.(3) Finally,the positive rate of the neuron-like cells was up to 73.5% in the group induced by ATRA with ACM.CONCLUSION: The ESCs are induced into neuron-like cells with high purity and efficiency by ATRA with ACM.

Aging changes of somatostatin (SS) mRNA-positive neurons in the dorsal hippocampus of old rats were studied by in situ hybridization histochemistry and image analysis. In the young rats,SS mRNA-positive cell bodies were mainly located in the pyramidal and polymorphic cell layers of the CA1 and CA2 regions, and in the strata radiatum and polymorphic cell layer of the CA3 region, and in the polymorphic cell layer of dentate gyrus. In the old rats,SS mRNA-positive cell bodies were concentrated in the polymorphic cell layer and denate gyrus. A marked decrease of number of SS mRNA-positive cell bodies and a significant increase of gray value of SS mRNA-positive cell bodies were found in the dorsal hippocampus and dentate gyrus of the old rats as compared with the young rats. No significant age difference in the sectional area of SS mRNA-positive cell bodies was observed in the dorsal hippocampus. The results showed that there were apparent changes with aging of the SS mRNA-positive cell bodies in the dorsal hippocampus of the rats. The significance of the changes remains to be studied further.

Objective To investigate the application of auricular cartilage in nasal plastic surgery and its clinical effects of the different tip graft and reconstructed domal graft (RDG).Methods From October 2009 to May 2015,the surgical procedure was performed on 286 patients,through the application of cymba and conchae cartilages into various tip graft and RDG in nasal tip plasty.Results A total of 286 patients were followed up post-operatively for 3 months to 5 years,245 patients (85.7％ achieved expected stability tip,the tip down to 1-2 mm after one year.Tip skin thinning was observed in 10 patients after one year,nasal tip down occurred in 89 patients;the columella skew and asymmetric nostrils occurred in 31 patients.Conclusions The ear cartilage can be made into various tip graft for rhinoplasty,with satisfactory clinical outcomes.

Objective To demonstrate the carotid artery hemodynamic characteristics in patients with common carotid artery occlusion,and to investigate the value of color Doppler ultrasonography in evaluating it.Methods 30 patients with common carotid artery (CCA) occlusion or severe stenosis (stenosis rate＞70％) were as the patients group,while 30 volunteers with the normal indicators of carotid artery ultrasonography as the control group.In both groups,the examinations of the CCA,the internal carotid artery (ICA),the external carotid artery (ECA) and the vertebral artery (VA) were taken by color Doppler ultrasonography under quiescent conditions.Bilateral carotid ultrasonography studies were performed,which included the peak systolic velocity (PSV),end-diastolic velocity (EDV),mean velocity (MV),and resistance index (RI).Results The peak systolic velocity (PSV),EDV and MV of ICA were obviously lower in case group at affected side than in the control group (P＜0.05 or 0.01).The blood flow of ECA at the affected side was retrograde or bidirectional.PSV,EDV and MV of the external carotid artery (ECA) were obviously higher in the healthy side in patients than in the control group (all P＜0.01).There was a significant difference in blood flow velocity of bilateral vertebral artery (VA) between the patients group and the control group,and the PSV,EDV and MV were obviously higher in the former than in the latter (all P＜0.01).And the RI was different between the two groups (P＜0.05).Conclusions The pressure of ICA is dropped after the common carotid artery occlusion,and then ECA reversely feeds ICA by collateral circulation.The blood flow velocities of bilateral vertebral artery and contralateral ECA are significantly increased to compensate for cerebral blood supply through collateral circulation channels.Color Doppler ultrasonography can reflect this phenomenon more fully and accurately,and provide clinically valuable diagnostic information.

Objective To investigate the possibility and utility of metformin alone or in combination with fosinopril to reduce blood pressure in patients with essential hypertension.Met hods A total of 140 cases of non-diabetic essential hypertension with hyperinsulinemia were recruited and randomly assigned to two groups: a group of 68 treated with metformin 500 mg tid and a group of 72 treated with fosinopril 10 mg qd.The duration of the treatment was 8 weeks.Combination therapy with the two drugs was used after 4 weeks of treatment if needed.If the target goals of systolic blood pressure (SBP) < 140 mm Hg (1 mm Hg =0.133 kPa) and /or diastolic blood pressure (DBP) <90 mm Hg were not attained 4 weeks, combination therapy with two drugs was used in either group in the next 4 weeks.The changes of blood pressure and insulin sensitivity of the two groups were observed before and after treatment.Results (1) After 4 weeks of treatment, SBP in metformin group and fosinopril group decreased by ( 13.0 ± 1.2) mm Hg and (15.4 ± 1.4) mm Hg, and DBP decreased by (9.0 ± 1.0) mm Hg and ( 10.4 ± 1.1 ) mm Hg respectively.After 8 weeks of treatment, SBP in metformin group and fosinopril group decreased by (17.8 ± 1.5) mm Hg and (20.9 ± 1.5) mm Hg, and DBP decreased by (13.2 ±0.9) mm Hg and (15.3 ± 1.1) mm Hg respectively.There was no significant difference in the decline of blood pressure between the two groups (P >0.05).The rates of combination therapy were both 54% in the two groups.(2) Fasting insulin as well as 30 min and 120 min insulin levels after oral glucose tolerance test and insulin area under the curve in the metformin group were significantly reduced after 4 and 8 weeks of treatment as compared with those of baseline(P < 0.05 and P < 0.01 ) .In the fosinopril group, however, they decreased only after 8 weeks treatment (P < 0.05).The insulin action index in the metformin group was higher than that in the fosinopril group after 4 weeks of treatment (P <0.05) ,but there was no significant difference between the two groups after 8 weeks of treatment (P > 0.05).Conclusion Metformin and fosinopril have similar antihypertensive effect and a good synergy in essential hypertension with hyperinsulinemia.

Objective To prepare natural anti-keratin IgM monoclonal antibody. Methods Spleen cells of BALB/c mice raised in specific pathogen free conditions were directly fused with Sp2/0 myeloma cells. The hybridoma supernatants were tested by ELISA using pre-extracted keratin. The natural IgM obtained was further identified by immunochemistry and immunoblot methods. Results The cell fusion rate was about 60% without pre-immunization. About 14% supernatants reacted with the keratin antigen. Three hybridoma strains secreting natural IgM monoclonal antibody against keratin were obtained. The immunochemistry results showed that the natural anti-keratin IgM was able to bind to epidermis, sebaceous gland, hair follicule, and muscle tissues. Conclusion B lymphocytes in normal BALB/c mice spleen can produce natural antibody against kerain.

Objectives To investigate the binding of natural IgM to Staphylococcus aureus and its role in the phagocytosis of S. aureus by phagocytes, and to pave way for further study on the role and mechanism of natural IgM in defense of bacteria. Methods The binding of natural antikeratin IgM 3B4 to S. aureus was analyzed by ELISA and indirect immunoiluorescence. The role of 3B4 in the phagocytosis was analyzed by colony forming assay and flow cytometry (FCM). Results Both ELISA and indirect immunofluorescence proved the binding of natural IgM 3B4 to S. aureus. Colony forming assay found that the amount of colony forming units decreased significantly when 3B4 was added. The analysis of FCM showed that 3B4 augmented phagocytosis of 5. aureus by phagocytes. Conclusions Natural antikeratin IgM 3B4 can bind to S. aureus and regulate the phagocytosis of it, indicating that natural IgM may play some role in the defense against bacterial infection.

AIM:To explore the supportive and expansive effects of aorta-gonad-mesonephros(AGM) region derived stromal cells on hematopoietic stem cells(HSC) in vitro.METHODS:The murine stromal cells were separated and cultured from AGM region of a 11 day postcoitum(dpc) mouse embryo and 6 week mouse.After identification by Wright's staining and flow cytometry,the stromal cells were co-cultured with the embryonic stem cell(ESC)-derived,cytokine-induced HSCs,and the maintenance and expansion of HSCs were evaluated by detecting CD34+,CD34+Sca-1+cells with flow cytometry.Blast colony-forming cell(BL-CFC) and high proliferative potential colony-forming cells(HPP-CFC) were determined by semi-solid medium clonal culture.RESULTS:AGM-derived and bone marrow(BM)-derived stromal cells were similar in morphology and phenotype,and had common character of stromal cells.Supported by AGM stromal cells or by BM stromal cells,more primitive progenitor cells HPP-CFC were expanded,but BL-CFC expansion was only detected in AGM-derived stromal cells.In the supporting of BM stromal cells CD34+ hematopoietic stem/progenitor cells were expanded 3-4 times,but no significant expansion in CD34+Sca-1+ cells was observed.While in the supporting of AGM stromal cells,both CD34+ hematopoietic stem/progenitor cells and CD34+Sca-1+ cells were expanded significantly from 4 to 5 times,respectively(P

Objective To investigate UV- or heat-inactivated Epstein-Barr virus(EBV)stimulating the production of anti-keratin autoantibody(AK auto Ab)in cultured human umbilical cord blood B cells. Methods Mononuclear cells were isolated routinely from umbilical cord blood, in which monocytes, NK cells and cytotoxicity T cells were eliminated by L-leucine methyl ester method, and T cells were removed by sheep red blood cells(SRBCs)treated with 2-amino ethyl-isothiouronium bromide (AET). The purified B cells were treated with UV- or heat-inactivated EBV respectively and then cultured in complete IMDM. CD5, CD3, CD4 and CD8 cells were detected by flow cytometry. IgG and IgM of AK auto Ab were measured by ELISA in the supernatant which came from the B cells treated by UV-inactivated EBV or EBV-transformed B cells respectively. Results In UV-inactivated EBV group CD5+B cells accounted for 43% and 47% of all cells detected on the 14th and 28th day, respectively. No CD3, CD4 and CD8 cells were detected during this period. In UV-inactivated EBV group the AK auto Ab of IgG and IgM increased significantly on the 18th and 26th day, respectively (P 0.05). On the 40th day the AK auto Ab of IgG and IgM were significantly higher in EBV-transformed B cell group than those in UV-inactivated EBV group. Conclusions UV-inactivated EBV is able to induce AK auto Ab production but heat-inactivated EBV does not, which suggests that EBV protein might be the effective agent in inducing the production of AK auto Ab.